4-methyldihydropyrimidinone compounds and pharmaceutical use thereof

ABSTRACT

The present invention relates to 4-methyldihydropyrimidinone compounds, or pharmaceutically acceptable salts thereof, having RORγ antagonist activity, pharmaceutical compositions comprising the same, and pharmaceutical use thereof. A compound of Formula (1) or (2) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising the same, and pharmaceutical use thereof are provided.

TECHNICAL FIELD

The present invention relates to 4-methyldihydropyrimidinone compounds,or pharmaceutically acceptable salts thereof, having RORγ antagonistactivity, pharmaceutical compositions comprising the same, andpharmaceutical use thereof.

BACKGROUND ART

RORγ (i.e., Retinoid-related Orphan Receptor gamma) is a nuclearreceptor which is important for the differentiation and activation ofTh17 cells. RORγt is also known as a splicing variant of RORγ (Nonpatent literature 1). RORγ and RORγt differ only in their N-terminaldomains and share the same ligand-binding domain and DNA-binding domain.It is reported that RORγ is expressed in other tissues besides Th17cells (Non Patent Literature 1).

Inhibition of RORγ can inhibit the differentiation and activation ofTh17 cells. IL-17 produced in Th17 cells is involved in the induction ofa variety of chemokines, cytokines, metalloproteases, and otherinflammatory mediators and the migration of neutrophil, and therefore,inhibition of IL-17 may lead to inhibit such induction and migration(Non Patent Literatures 2 and 3). It is known that. Th17 cells areinvolved in autoimmune diseases (such as rheumatoid arthritis,psoriasis, inflammatory bowel disease (such as Crohn's disease andulcerative colitis), multiple sclerosis, systemic lupus erythematosus(SLE), Behcet's disease, sarcoidosis, Harada disease, ankylosingspondylitis, uveitis, polymyalgia rheumatics, type I diabetes,graft-versus-host disease, alopecia areata, and vitiligo), allergicdiseases, dry eye, fibrosis (such as lung fibrosis and primary biliarycirrhosis), and cancers (such as malignant melanoma and prostatecancer).

RORγ in adipose tissues is related to the regulation of adipogenesis andinhibition of RORγ can ameliorate insulin resistance (Non PatentLiterature 4). It is known that adipose tissues are involved inmetabolic diseases (such as hepatic steatosis).

It is also known that 11-17 and Th17 cells are involved in ischemia,cardiomyopathy, hypertension, and periodontitis.

For example, as for rheumatoid arthritis, it is reported thatadministration of anti-IL-17 antibody can ameliorate swelling and jointdestruction associated with collagen-induced arthritis (Non PatentLiterature 5). It is also reported that swelling and joint destructionassociated with collagen-induced arthritis can be ameliorated inexperiments using IL-17-deficient mice (Non Patent Literature 6).

As for psoriasis, it is reported that administration of anti-TL-17antibody is effective in treating psoriasis in clinical trials (NonPatent Literature 7). Anti IL-17 antibodies have been placed on themarket for use in psoriasis (Nor Patent Literature 8).

As for inflammatory bowel diseases such as Crohn's disease andulcerative colitis, adaptive transfer of T cells derived from RORγ-KOmice does not increase IL-17 in the mucosa in a colitis model induced bythe adaptive transfer of T cells, thereby the onset of colitis can beinhibited (Non Patent Literature 9) is also reported that an anti-IL-23antibody, an antibody against 11-23 which activates Th17 cell s, iseffective in treating Crohn's disease in clinical trials (Non PatentLiterature 20).

As for multiple sclerosis, the disease state of a mouse experimentalautoimmune encephalomyelitis model which is an animal model of multiplesclerosis can be inhibited in RORγ-KO mice (Non Patent Literature 10).It is also reported that an anti-IL-17A antibody can ameliorate MRIobservation in relapsing remitting multiple sclerosis in clinical trials(Non Patent Literature 21).

As for systemic lupus erythematosus, it is reported that administrationof anti-IL-17 antibody can inhibit onset of GEM nephritis model inRORγt-KO mice which is an animal model of glomerulonephritis (Non PatentLiterature 11). Administration of anti-IL-17 antibody potentiallyinhibits nephritis associated with SLE as well. (Non Patent Literature12).

As for ankylosing spondylitis, it is reported that administration ofanti-IL-17 antibody is effective in treating ankylosing spondylitis (NonPatent Literature 13).

As for uveitis, it is reported that administration of anti-IL 17antibody is effective in treating uveitis associated with Behcet'sdisease, sarcoidosis, and Harada disease (Non Patent Literature 7).

As for polymyalgia rheumatica, efficacy of anti-IL-17 antibody iscurrently assessed in clinical trials for polymyalgia rheumatica.

As for type I diabetes, administration of anti-IL-17 antibody caninhibit progression of disease states in a NOD mouse model which is atype I diabetes model (Non Patent Literature 14). Efficacy ofanti-IL-17A antibody is currently assessed in clinical trials (NonPatent Literature 22).

As for graft-versus-host disease, it is reported that transfection ofRORγ-KO-mouse-derived cells can ameliorate survival rates and rejectionsin a host in a mouse transplant model (Non Patent Literature 19).

As for alopecia areata, efficacy of anti-IL-17A antibody is currentlyassessed in clinical trials (Non Patent Literature 25).

As for vitiligo, increases of IL-17 and Th17 cells are recognized inpatient sera and pathological tissues, respectively (Non PatentLiterature 39).

As for allergic diseases such as asthma, attenuated eosinophilicpulmonary inflammation, the reduced number or CD4+ lymphocytes, and thedecrease of Th2 cytokines/chemokines levels are exhibited RORγ-KO micein an OVA-sensitized model, which then allergic reactions can beinhibited (Non Patent Literature 15). Efficacy of anti-IL-17A antibody scurrently assessed in clinical trials for atopic dermatitis (Non PatentLiterature 23). Efficacy of anti-IL-23 antibody is currently assessed inclinical trials for asthma (Non Patent Literature 24).

As for dry eye, it is reported that Th17 cells increase in an animalmodel of dry eye, and efficacy of anti-IL-17 antibody is currentlyassessed in clinical trials for dry eye patients (Non Patent Literature16).

As for fibrosis, administration of anti-IL-17 antibody can inhibitinflammation and fibrosis in lung and extend survival of animals in ableomycin-induced lung fibrosis model which is an animal model of lungfibrosis (Non Patent Literature 17).

As for primary biliary cirrhosis, it is reported that Th17 cellsincrease in the lesion area of patients with primary biliary cirrhosis,and efficacy of anti-IL-23 antibody is currently assessed in clinicaltrials (Non Patent Literature 18).

As for malignant melanoma, efficacy of anti-IL-17 antibody is currentlyassessed in clinical trials (Non Patent Literatures 26 and 27).

As for prostate cancer, it is recognized that anti-IL-antibody treatmentdecreased the formation of micro-invasive prostate cancer in Pten-nullmice (Non Patent Literature 33).

As for insulin resistance, the insulin resistance induced by feedinghigh-fat diets can be inhibited in RORγ KO mice (Non Patent Literature4).

As for hepatic steatosis, it is recognized that anti-IL-17 antibodyameliorated steatosis on pathological tissues in an alcoholicliver-disease model (Non Patent Literature 34).

As for non-alcoholic fatty liver disease, lit is recognized thatanti-IL-17 antibody treatment improved liver function, attenuatedhepatic lipid accumulation, suppressed Kupffer cells activation, anddecreased proInflammatory cytokines levels in a high fat diet-inducednon-alcoholic fatty liver disease model (Non Patent Literature 35).

As for ischemia and cardiomyopathy, it is reported that IL-17Acontributes to myocardial ischemia/reperfusion injury by regulatingcardio myocyte apoptosis and neutrophil infiltration. It is recognizedthat anti-IL-17A antibody treatment or IL-17A knockout reduced infarctsize, improved cardiac function, and thus, amelioratedischemia/reperfusion injury (Non Patent Literature 36).

As for hypertension, it is reported that treatment with antibody againstIL-17A or IL-17RA suppressed increased blood pressure by administrationof angiotensin II (Non Patent Literature 37).

As for periodontitis, increase of Th17 cells or IL-17 was recognized inan experimental periodontitis model is reported that treatment with RORγantagonist, GSK805, or anti-IL-17A antibody diminished bone loss in themodel (Non Patent Literature 38).

On the basis of these findings, RORγ antagonists are deemed to bebeneficial for preventing or treating autoimmune diseases, allergicdiseases, dry eye, fibrosis, cancers (such as malignant melanoma andprostate cancer), metabolic disease, ischemia, cardiomyopathy,hypertension, and periodontal disease.

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SUMMARY OF INVENTION

The present invention provides 4-methyldihydropyrimidinone compounds, orpharmaceutically acceptable salts thereof, having RORγ antagonistactivity, pharmaceutical compositions comprising the same, and theirmedical use. One aspect of the present invention includes the followingillustrative embodiments.

[Item 1]

A compound of Formula (1) or (2):

or a pharmaceutically acceptable salt thereof.

[Item 2]

The compound according to Item 1, wherein the compound is a compound ofFormula (1):

or a pharmaceutically acceptable salt thereof.

[Item 3]

The compound according to Item 1, wherein the compound is a compound ofFormula (2):

or a pharmaceutically acceptable salt thereof.

[Item 4]

A pharmaceutical composition comprising a compound according to any oneof Items 1 to 3 or a pharmaceutically acceptable salt thereof and apharmaceutically acceptable carrier.

[Item 5]

An RORγ antagonist comprising a compound according to any one of Items 1to 3 or a pharmaceutically acceptable salt thereof.

[Item 6]

A therapeutic or preventive agent for a disease selected from the groupconsisting of autoimmune diseases, allergic diseases, dry eye, cancers,metabolic disease, ischemia, cardiomyopathy, hypertension, andperiodontal disease, comprising a compound according to any one of Items1 to 3 or a pharmaceutically acceptable salt thereof.

[Item 7]

A method of antagonizing RORγ, comprising administering atherapeutically effective amount of compound according to any one ofItems 1 to 3 or a pharmaceutically acceptable salt thereof to a mammal.

[Item 8]

A method of treating or preventing a disease selected from the groupconsisting of autoimmune diseases, allergic diseases, dry eye, fibrosis,cancers, metabolic disease, ischemia, cardiomyopathy, hypertension, andperiodontal disease, comprising administering a therapeuticallyeffective amount of a compound according to any one of Items 1 to 3 or apharmaceutically acceptable salt thereof to a mammal.

[Item 9]

Use of a compound according to any one of Items 1 to 3 or apharmaceutically acceptable salt thereof the manufacture of an RORγantagonist.

[Item 10]

Use of a compound according to any one of Items 1 to 3 or apharmaceutically acceptable salt thereof in the manufacture of atherapeutic or preventive agent for a disease selected from the groupconsisting of autoimmune diseases, allergic diseases, dry eye, fibrosis,cancers, metabolic disease, ischemia, cardiomyopathy, hypertension, andperiodontal disease.

[Item 11]

A compound according to any one of Items 1 to 3 or a pharmaceuticallyacceptable salt thereof for use in an RORγ antagonist.

[Item 12]

A compound according to any one of Items 1 to 3 or a pharmaceuticallyacceptable salt thereof for use in treating or preventing a diseaseselected from the group consisting of autoimmune diseases, allergicdiseases, dry eye, fibrosis, cancers, metabolic disease, ischemia,cardiomyopathy, hypertension, and periodontal disease.

[Item 13]

A commercial package comprising a pharmaceutical composition accordingto Item 4 and a package insert concerning the pharmaceutical compositiondescribing that the pharmaceutical composition can be used for treatingor preventing a disease selected from the group consisting of autoimmunediseases, allergic diseases, dry eye, fibrosis, cancers, metabolicdisease, ischemia, cardiomyopathy, hypertension, and periodontaldisease.

[Item 14]

A kit comprising a pharmaceutical composition according to Item 4 and apackage insert concerning the pharmaceutical composition describing thatthe pharmaceutical composition can be used for treating or is preventinga disease selected from the group consisting of autoimmune diseases,allergic diseases, dry eye, fibrosis, cancers, metabolic disease,ischemia, cardiomyopathy, hypertension, and periodontal disease.

[Item 15]

A crystalline form of a compound of Formula (1):

showing a powder X-ray diffraction pattern having any three or morepeaks selected from the group consisting of 7.4±0.2°, 9.9±0.2°,10.5±0.2°, 11.4±0.2°, 11.6±0.2°, 13.4±0.2°, 14.2±0.2°, 17.4±0.2°,18.3±0.2°, 18.7±0.2°, and 19.4±0.2° of the diffraction angle (2θ)measured with CuKα radiation.

[Item 16]

A monohydrate of a compound of Formula (1):

[Item 17]

A crystalline form of a monohydrate of a compound of Formula (1):

showing a powder X-ray diffraction pattern having any three or morepeaks selected from the group consisting of 4.2±0.2°, 9.7±0.2°,13.7±0.2°, 14.0±0.2°, 15.2±0.2°, 15.4±0.2°, 16.9±0.2°, 18.8±0.2°,20.5±0.2°, 21.9±0.2°, and 22.4±0.2° of the diffraction angle (2θ)measured with CuKα radiation.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows a powder X-ray diffraction pattern of Crystalline Form A.The vertical axis shows diffraction intensity (cps: counts per second),and the horizontal axis shows diffraction angle 2θ (° C.).

FIG. 2 shows a differential scanning calorimetry (DSC) curve ofCrystalline Form A. The vertical axis shows Heat Flow (Watt per gram),and the horizontal axis shows temperature (° C.).

FIG. 3 shows a powder X-ray diffraction pattern of Crystalline Form B.The vertical axis shows diffraction intensity (cps: counts per second),and the horizontal axis shows diffraction angle 2θ (°).

FIG. 4 shows a DSC curve of Crystalline Form B. The vertical axis showsHeat Flow (Watt per gram), and the horizontal axis shows temperature (°C.).

FIG. 5 shows a powder X-ray diffraction pattern of Crystalline Form C.The vertical axis shows diffraction intensity (cps: counts per second),and the horizontal axis shows diffraction angle 2θ (°).

FIG. 6 shows a DSC curve of Crystalline Form C. The vertical axis showsHeat Flow (Watt per gram), and the horizontal axis shows temperature (°C.).

FIG. 7 shows a powder X-ray diffraction pattern of Crystalline Form D.The vertical axis shows diffraction intensity (cps: counts per second),and the horizontal axis shows diffraction angle 2θ (°).

FIG. 8 shows a TG-DTA curve of Crystalline Form D. The upper part ofvertical axis shows weight (gram), the lower part of vertical axis showstemperature (° C.), and the horizontal axis shows temperature (° C.).

FIG. 9 shows a DSC curve of Crystalline Form D. The vertical axis showsHeat Flow (Watt per gram), and the horizontal axis shows temperature (°C.).

FIG. 10 shows a powder X-ray diffraction pattern of Crystalline Form E.The vertical axis shows diffraction intensity (cps: counts per second),and the horizontal axis shows diffraction angle 2θ (0).

FIG. 11 shows a TG-DTA curve of Crystalline Form E. The upper part ofvertical axis shows weight (gram), the lower part of vertical axis showstemperature (° C.), and the horizontal axis shows temperature (° C.).

FIG. 12 shows a DSC curve of Crystalline Form E. The vertical axis showsHeat Flow (Watt per gram), and the horizontal axis shows temperature (°C.).

DESCRIPTION OF EMBODIMENTS

Definitions of terms used herein are shown as follows.

“A compound of Formula (1)” and “a compound of Formula (2)” are alsooptionally referred to as “Compound (1)” and “Compound (2)”,respectively. “Compound (1) or Compound (2)(,) or a pharmaceuticallyacceptable salt thereof” means Compound (1) or Compound (2) or apharmaceutically acceptable salt of Compound (1) or Compound (2), and isintended to include any of pharmaceutically acceptable salts of Compound(1) and pharmaceutically acceptable salts of Compound (2).

The term “pharmaceutically acceptable salt” may be any salts withoutexcess toxicity known in the art. Specifically, it includes, forexample, salts with inorganic acids, salts with organic acids, saltswith inorganic bases, and salts with organic bases. Various forms ofpharmaceutically acceptable salts are well known in the art and arelisted, for example, in the following references:

(a) Berge et al., J. Pharm. Sci., 66, p 1-19 (1977);

(b) Stahl et al., “Handbook of Pharmaceutical Salts: Properties,Selection, and Use” (Wiley-VCH, Weinheim, Germany, 2002);

(c) Paulekuhn et al., J. Med. Chem., 50, p 6665-6672 (2007).

According to known methods, Compound (1) or Compound (2) may be reactedwith ah inorganic acid, organic acid, inorganic base, or organic base togive each pharmaceutically acceptable salt thereof.

Such salts with inorganic acids include, for example, salts withhydrofluoric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid,nitric acid, phosphoric acid, and sulfuric acid. Preferable saltsinclude salts with hydrochloric acid, nitric acid, sulfuric acid,phosphoric acid, and hydrobromic acid.

Such salts with organic acids include, for example, salts with aceticacid, adipic acid, alginic acid, 4-aminosalicylic acid,anhydromethylenecitric acid, benzoic acid, benzenesulfonic acid, camphoracid, camphor-10-sulfonic acid, carbonic acid, citric acid, edetic acid,ethane-1,2-disulfonic acid, dodecylsulfonic acid, ethanesulfonic acid,fumaric acid, glucoheptonic acid, gluconic acid, glut uronic acid,glucoheptonic acid, glycoloylarsanilic acid, hydroxynaphthoic acid,2-hydroxy-1-ethanesulfonic acid, lactic acid, lactobionic acid, malicacid, maleic acid, mandelic acid, methanesulfonic acid, methylsulfuricacid, methylnitric acid, methylenebis(salicylic acid), galactaric acid,naphthalene-2-sulfonic acid, 2-naphthoic acid, 1,5-naphthalenedisulfonicacid, oleic acid, oxalic acid, pamoic acid, pantothenic acid, pecticacid, picric acid, propionic acid, polygalacturonic acid, salicylicacid, stearin acid, succinic acid, tannic acid, tartaric acid, teoclicacid, thiocyanic acid, trifluoroacetic acid, p-toluenesulfonic acid,undecanoic acid, asparaginic acid, and glutamic acid. Preferable saltsinclude salts with oxalic acid, maleic acid, citric acid, fumaric acid,lactic acid, malic acid, succinic acid, tartaric acid, acetic acid,trifluoroacetic acid, benzoic acid, glucuronic acid, oleic acid, pamoicacid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid,and 2-hydroxy-1-ethanesulfonic acid.

Such salts with inorganic bases include, for example, salts withlithium, sodium, potassium, magnesium, calcium, barium, aluminum, zinc,bismuth, and ammonium. Preferable salts include salts with sodium,potassium, calcium, magnesium, and zinc.

Such salts with organic bases include, for example, salts witharecoline, betaine, choline, clemizole, ethylenediamine,N-methylglucamine, N-benzylphenethylamine,tris(hydroxymethyl)methylamine, arginine, and lysine. Preferable saltsinclude salts with tris(hydroxymethyl)methylamine, N-methylglucamine,and lysine.

Compound (1) or Compound (2) or a pharmaceutically acceptable saltthereof may exist in a solvate form.

The term “solvate” means Compound (1) or Compound (2) or apharmaceutically acceptable salt thereof coordinate with a solventmolecule and includes a hydrate. Such a solvate is preferably apharmaceutically acceptable solvate and includes hydrates, ethanolates,and solvates with dimethylsulfoxide of Compound (1) or Compound (2) or apharmaceutically acceptable salt thereof.

Specifically, such a solvate includes a hemihydrate, monohydrate,dihydrate, or monoethanolate of Compound (1) or Compound (2), or amonohydrate of a hydrochloride salt or a 2/3 ethanolate of adihydrochloride salt of Compound (1) or Compound (2). A preferablesolvate includes a monohydrate of Compound (1). Such a solvate may beobtained according to known methods.

Compound (1) or Compound (2) or a pharmaceutically acceptable saltthereof may be labelled with an isotope atom such as ²H, ³H, ¹⁴C, and³⁵S.

For example, any hydrogen atoms of Compound (1) or Compound (2) includeprotium ¹H (H), deuterium ²H (D), and tritium ³H (T).

Compound (1) or Compound (2), or a pharmaceutically acceptable saltthereof is preferably Compound (1) or Compound (2), or apharmaceutically acceptable salt thereof, substantially purified. Morepreferable one is Compound (1) or Compound (2), or a pharmaceuticallyacceptable salt thereof, having 80% or more of purity.

A preferable crystalline form of Compound (1) or Compound (2), or apharmaceutically acceptable salt thereof, includes a crystalline form ofCompound (1) showing a powder X-ray diffraction pattern having at least3 peaks, for example, at least 3, 4, or 5 peaks, at any of 7.4±0.2°,9.9±0.2°, 10.5±0.2°, 11.4±0.2°, 11.6±0.2°, 13.4±0.2°, 14.2±0.2°,17.4±0.2°, 18.3±0.2°, 18.7±0.2°, or 19.4±0.2° of the diffraction angle(2θ) measured with CuKα radiation. A more preferable crystalline form ofCompound (1) may show a powder X-ray diffraction pattern having peaks at7.4±0.2°, 9.9±0.2°, and 13.4±0.2° of 2θ. A further preferablecrystalline form of Compound (1) may show a powder X-ray diffractionpattern having peaks at 7.4±8 0.2°, 9.9±0.2°, 13.4±0.2°, 18.7±0.2°, and19.4±0.2° of 2θ.

Another preferable crystalline form of Compound (1) or Compound (2), ora pharmaceutically acceptable salt thereof, includes a crystalline formof a monohydrate of Compound (1) showing a powder X-ray diffractionpattern having at least 3 peaks, for example, at least 3, 4, or 5 peaks,at any of 4.2±0.2°, 9.7±0.2°, 13.7±0.2°, 14.0±0.2°, 15.2±0.2°,15.4±0.2°, 16.9±0.2°, 18.8±0.2°, 20.5±0.2°, 21.9±0.2°, or 22.4±0.2° ofthe diffraction angle (20) measured with CuKα radiation. A morepreferable crystalline form of a monohydrate of Compound (1) may show apowder X-ray diffraction pattern having peaks at 4.2±0.2°, 9.7±0.2°, and16.9±0.2° of 20. A further preferable crystalline form of a monohydrateof Compound (1) may show a powder X-ray diffraction pattern having peaksat 4.2±0.2°, 9.7 0.2°, 13.7±0.2°, 15.2±0.2°, and 16.9±0.2 of 20.

The error range of the diffraction angle (2θ) in a powder X-raydiffraction pattern is preferably ±0.2°, more preferably ±0.1°, andfurther preferably ±0.05°.

According to known methods in the art of pharmaceutical formulations, apharmaceutical composition herein may be prepared by, for example,mixing Compound (1) or Compound (2) or a pharmaceutically acceptablesalt thereof with at least one or more pharmaceutically acceptablecarrier(s) in an appropriate amount. The content (also referred to as “atherapeutically effective amount” herein) Compound (1) or Compound (2)or a pharmaceutically acceptable gait thereof in the pharmaceuticalcomposition varies depending on dosage forms and doses and is, forexample, 0.1 to 100% by weight of the composition.

A dosage form of Compound (1) or Compound (2) or pharmaceuticallyacceptable salt thereof includes an oral preparation such as tablets,capsules, granules, powders, lozenges, syrups, emulsions, andsuspensions and a parenteral preparation such as external preparations,suppositories, injections, eye drops, nasal preparations, and pulmonarypreparations.

The term “pharmaceutically acceptable carrier” includes variousconventional organic or inorganic carrier substances for formulationmaterials such as excipients, disintegrants, binders, fluidizers, andlubricants in solid formulations; solvents, solubilizing agents,suspending agents, tonicity agents, buffers, and soothing agents inliquid formulations; and bases, emulsifying agents, wetting agents,stabilizers, stabilizing agents, dispersants, plasticizers, pHregulators, absorption promoters, gelators, preservatives, fillers,solubilizers, solubilizing agents, and suspending agents semisolidformulations. A preserving agent, an antioxidant agent, a colorant, or asweetening agent may also be optionally used as an additive.

Such an “excipient” includes, for example, lactose, white soft sugar,D-mannitol, D-sorbitol, cornstarch, dextrin, microcrystalline cellulose,crystalline cellulose, carmellose, carmellose calcium, sodiumcarboxymethyl starch, low substituted hydroxypropyl cellulose, and gumarabic.

Such a “disintegrant” includes, for example, carmellose, carmellosecalcium, carmellose sodium, sodium carboxymethyl starch, croscarmellosesodium, crospovidone, low substituted hydroxypropyl cellulose,hydroxypropyl methylcellulose, and crystalline cellulose.

Such a “hinder” includes, for example, hydroxypropyl cellulose,hydroxypropyl methylcellulose, povidone, crystalline cellulose, whitesoft sugar, dextrin, starch, gelatin, carmellose sodium, and gum arabic.

Such a “fluidizer” includes, for example, light anhydrous silicic acidand magnesium stearate.

Such a “lubricant” includes, for example, magnesium stearate, calciumstearate, and talc.

Such a “solvent” includes, for example, purified water, ethanol,propyleneglycol, macrogol, sesame oil, corn oil, and olive oil.

Such a “solubilizing agent” includes, for example, propyleneglycol,D-mannitol, benzyl benzoate, ethanol, triethanolamine, sodium carbonate,and sodium citrate.

Such a “suspending agent” includes, for example, benzalkonium chloride,carmellose, hydroxypropyl cellulose, propyleneglycol, povidone,methylcellulose, and glyceryl monostearate.

Such a “tonicity agent” includes, for example, glucose, D-sorbitol,sodium chloride, and D-mannitol.

Such a “buffer” includes, for example, sodium hydrogen phosphate, sodiumacetate, sodium carbonate, and sodium citrate.

Such a “soothing agent” includes, for example, benzyl alcohol.

Such a “base” includes, for example, water, animal or vegetable oilssuch as olive oil, corn oil, arachis oil, sesame oil, and castor oil,lower alcohols such as ethanol, propanol, propylene glycol, 1,3-butyleneglycol, and phenol, higher fatty acids and esters thereof, waxes, higheralcohols, polyalcohols, hydrocarbons such as white petrolatum, liquidparaffin, and paraffin, hydrophilic petrolatum, purified lanolin,absorptive ointment, hydrous lanolin, hydrophilic ointment, starch,pullulan, gum arabic, tragacanth gum, gelatin, dextran, cellulosederivatives such as methylcellulose, carboxymethylcellulose,hydroxyethyl cellulose, and hydroxypropyl cellulose, synthetic polymerssuch as carboxyvinyl polymers, sodium polyacrylate, polyvinyl alcohol,and polyvinylpyrrolidone, propylene glycol, macrogol such as macrogol200 to 600, and a combination of any two or more of them.

Such a “preserving agent” includes, for example, ethylparahydroxybenzoate, chlorobutanol, benzyl alcohol, sodiumdehydroacetate, and sorbic acid.

Such an “antioxidant agent” includes, for example, sodium sulfite andascorbic acid.

Such a “colorant” includes, for example, food dye such as Food Red No. 2and No. 3, and Food Yellow No. 4 and No. 5, and β-carotene.

Such a “sweetening agent” includes, example saccharin sodium,dipotassium glycyrrhizate, and aspartame.

A pharmaceutical composition herein may be administered orally orparenterally such as locally, rectally, intravenously, intramuscularly,and subcutaneously to mammals other than human such as mice, rats,hamsters, guinea pigs, rabbits, cats, dogs, pigs, cattle, horses, sheep,and monkeys as well as human dose may vary depending on subjects to beadministered, diseases, symptoms, dosage forms, routes ofadministration, etc. For example, in oral administration to an adultpatient, the dose of Compound (1) or Compound (2), the activeingredient, ranges generally from about 0.01 mg to about 1 g per day,which may be administered once or several times in a divided amount.

A kit such as kits for administration, treatment, and/or prevention, apackage such as packaged goods, and a set and/or case of medicine whichcomprises a pharmaceutical composition comprising Compound (1) Compound(2) or a pharmaceutically acceptable salt thereof as the activeingredient or active agent and a written matter concerning thecomposition indicating that the composition may or should be used fortreatment and/or prevention are also useful. Such a kit, package, andset of medicine may comprise one or more containers filled with thepharmaceutical composition or one or more active ingredients and otherdrugs or medicines (or ingredients) used for the composition. Examplesof such a kit, package, and set of medicine include commercial kits,commercial packages, and commercial medicine set for appropriate use inthe treatment and/or prevention of intended diseases. The written mattercomprised in such a kit, package, and set of medicine includes acautionary note or package insert in the form designated by thegovernment organization that regulates manufactures, use, or sales ofpharmaceutical or biological products which ensures an approval by thegovernment organization on manufactures, use, or sales of productsconcerning administration to humans. The kit, package, and set ofmedicine may include packaged products as well as structures configuredfor appropriate administration steps and configured so as to be able toachieve more preferable medical treatment and/or prevention includingtreatment and/or prevention of intended diseases.

Compound (1) or Compound (2) or a pharmaceutically acceptable saltthereof has RORγ antagonism and is useful for an RORγ antagonist.

The term “having RORγ antagonist activity”, “having RORγ antagonism”, or“antagonizing RORγ” means that the function of RORγ is antagonized,preferably specifically antagonized, to disappear or reduce itsactivity, and includes, for example, antagonizing, preferablyspecifically antagonizing, the function of RORγ according so theconditions described in Test Example 1 below.

The term “RORγ antagonist” means any substances that antagonize thefunction of RORγ, preferably any substances that specifically antagonizethe function of RORγ.

The term “RORγ” is preferably “human RORγ”.

Compound (1) or Compound (2) or a pharmaceutically acceptable saltthereof has RORγ antagonism, and is expected to be effective againstdiseases that involve the function of RORγ.

Specifically, Compound (1) or Compound (2) or a pharmaceuticallyacceptable salt thereof is expected to be useful for treating orpreventing a disease selected from the group consisting of autoimmunediseases, allergic diseases, dry eye, fibrosis, cancers, metabolicdisease, ischemia, cardiomyopathy, hypertension, and periodontaldisease.

The term “autoimmune diseases” means a generic name of diseases where animmune system of a subject overreacts to and attacks even normal cellsand tissues thereof to cause symptoms, and includes, specifically,rheumatoid arthritis, psoriasis, inflammatory bowel diseases such asCrohn's disease and ulcerative colitis, multiple sclerosis, systemiclupus erythematosus (SLE), Behcet's disease, sarcoidosis, Haradadisease, ankylosing spondylitis, uveitis, polymyalgia rheumatica, typediabetes, graft-versus-host disease, alopecia areata, and vitiligo.

The term “allergic diseases” means diseases derived from the conditionwhere an immune reaction excessively occurs against a certain antigen,and includes, specifically atopic dermatitis, allergic rhinitis such aspollen allergy, allergic conjunctivitis, allergic gastroenteritis,asthma such as bronchial asthma and infantile asthma, food allergy,medication allergy, and hives.

The term “fibrosis” means a condition with increased fibroconnectivetissues, and includes, specifically, lung fibrosis and primary biliarycirrhosis.

The term “cancers” includes, specifically, malignant melanoma andprostate cancer.

The term “metabolic disease” means a disease caused by abnormality ofmetabolic turnover or a disease which includes metabolic abnormality asan element that constitutes pathogenesis, and includes, for example,diabetes such as type diabetes and type II diabetes, hepatic steatosis,and non-alcoholic fatty liver disease.

The term “treating” used herein also includes ameliorating symptoms,preventing from becoming severe, maintaining remission, preventingexacerbation, and preventing relapse.

The term “preventing” used herein means suppressing pathogenesis ofsymptoms.

Compound (1) or Compound (2), or a pharmaceutically acceptable saltthereof, has the following demonstrated properties:

(i) high metabolic stability referring to Test Example 2;(ii) beneficial pharmacokinetic profile, including favorable plasmahalf-life referring to Test Example 5;(iii) low potency to induce drug metabolic enzymes such as CYP3A4referring to Test Example 3;(iv) favorably high solubility referring to Test Example 4; andv) sustained and/or potent pharmacological effect referring to TestExamples 6 and 7.

These properties render Compound (1) or Compound (2), or apharmaceutically acceptable salt thereof, particularly advantageous. Forexample, Compound (1) or Compound (2), or a pharmaceutically acceptablesalt thereof:

(i) exhibits sustained pharmacological effects that may allow for lowerdosing frequency or longer dosing intervals, which may favorably impactpatient compliance and thereby improve overall therapeutic outcome;(ii) exhibits a low induction of drug metabolic enzymes such as CYP3A4,which may result in decreased metabolism of concomitant drugsmetabolized by such enzymes, and thus the present compound may be moresuitable for patients simultaneously taking multiple therapeutic drugs;and(iii) may have a favorable oral bioavailability based upon highsolubility and, thus, may express dose-dependent increase in plasmaconcentration even in high dosing amount and/or exhibit small individualvariability in absorption process.

As long as an embodiment disclosed herein compatible with anotherembodiment disclosed in another portion of the description, any two ormore combinations of these embodiments are also intended to be includedin the invention.

A method of preparing Compound (1) or Compound (2) or a pharmaceuticallyacceptable salt thereof is illustrated in the examples as below. Amethod of preparing Compound (1) or Compound (2) or a pharmaceuticallyacceptable salt thereof, however, is not intended to be limited thereto.

Each compound obtained in each step may be isolated and/or purified byknown methods such as distillation, recrystallization, and columnchromatography, if necessary, but each reaction may optionally proceedto a sequential step without isolation and/or purification.

The room temperature herein means a temperature under no control, andincludes 1° C. to 40° C. as one embodiment.

EXAMPLES

¹H-NMR spectra were measured with tetramethylsilane as an internalstandard in CDCl₃ or DMSO-d₆ and all 5 values are shown in ppm. Symbolsin spectral data mean as follows.

s: singletd: doublett: tripletq: quartetdd: double doubletddd: double double doubletbrs: broad singletm: multipletJ: coupling constant

Example 1 Synthesis of3-{(S)-4-[4-((1R,2R)-2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid Step 1 (S)-2-Methyl-propane-2-sulfinic acid[1-(4-bromo-3-chloro-phenyl)-eth-(E)-yliden]-amide

1-(4-Bromo-3-chloro-phenyl)-ethanone (60.0 g) and(S)-(−)-2-methyl-propane-2-sulfinic acid amide (37.3 q) were mixed incyclopentylmethyl ether (257 mL). To the reaction solution was addedtetraethyl orthotitanate (70.3 g), and the reaction solution was stirredat 100° C. for 5 boats. To the reaction solution was added 40 w/v %aqueous ammonium lactate solution (308 mL) at room temperature, and themixture was extracted with ethyl acetate. The resulting organic layerwas washed with water and brine, and then dried over magnesium sulfate.Magnesium sulfate was removed through a filter, and then the filtratewas concentrated under reduced pressure. The resulting residue waspurified by, silica gel column chromatography (ethylacetate:n-hexane=1:4→2:3). The resulting residue was solidified withn-hey ether and the precipitated solid was filtered to give the titlecompound (70.6 g). ¹H-NMR (400 MHz, CDCl₃) 1.30 (s, 9H), 2.72 (s, 3H),7.58 (dd, J=8.55, 2.08 Hz, 1H), 7.66 (d, J=8.55 Hz, 1H), 7.91 (d, J=2.08Hz, 1H)

Step 2(R)-3-(4-Bromo-3-chloro-phenyl)-2-isopropyl-3-((S)-2-methyl-propane-2-sulfinylamino)butyricacid methyl ester

Diisopropylamine (12.4 mL) was mixed with tetrahydrofuran (89.1 mL)under argon. To the reaction solution was added dropwise a 2.66Mn-butyllithium/n-hexane solution (33.5 mL) at −78° C., and the reactionsolution was stirred for 5 minutes under ice cooling. To the reactionsolution was added, dropwise a mixed solution of 3-methyl-butyric acidmethyl ester (11.7 mL) in tetrahydrofuran (44.5 mL), and the reactionsolution was stirred at −78° C. for 1 hour. To the reaction solution wasadded dropwise a mixed solution of (S)-2-methyl-propane-2-sulfinic acid[1-(4-bromo-3-chloro-phenyl)-eth-(E)-yliden]-amide (15.0 g) intetrahydrofuran (44.5 mL), and she reaction solution was stirred for 2hours. To the reaction solution was added a mixed solution of aceticacid (5.1 mL) in to of (25.4 mL), and the mixture was stirred under roomtemperature. To the reaction solution was added 1M aqueous monosodiumcitrate solution (100 ml), and the layers were separated. The resultingorganic layer was washed with water (100 ml, twice). Combined aqueouslayers were extracted with ethyl acetate (100 mL, twice). Combinedorganic layers were washed with 1M aqueous monosodium citrate solution(100 ml), water (100 mL, twice), and brine, and then dried overmagnesium sulfate. Magnesium sulfate was removed through a filter, andthen the filtrate was concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography (ethylacetate:n-hexane=1:4→3:2) to give the title compound (1.8.0 g).

¹H-NMR (400 MHz, CDCl₃) 0.76 (d, J=6.94 Hz, 1.2H), 0.93 (d, J=6.94 Hz,1.8H), 0.95 (d, J=6.94 Hz, 1.8H), 1.00 (d, J=6.94 Hz, 1.2H), 1.26 (s,5.4H), 1.34 (s, 3.6H), 1.74-1.81 (m, 0.4H), 1.87 (s, 1.2H), 1.88 (s,1.8H), 2.04-2.13 (m, 0.6H), 2.46 (d, J=3.93 Hz, 0.4H), 2.80 (d, J=3.93Hz, 0.6H), 3.60 (s, 1.8H), 3.70 (s, 1.2H), 5.13 (s, 0.6H), 5.42 (s,0.4H), 7.17-7.21 (m, 1H), 7.54-7.59 (m 2H)

Step 3 (S)-2-Methyl-propane-2-sulfinic acid[(R)-1-(4-bromo-3-chloro-phenyl)-2-hydroxymethyl-1,3-dimethyl-butyl)amide

(R)-3-(4-Bromo-3-chloro-phenyl)-2-isopropyl-3-((S)-2-methyl-propane-2-sulfinylamino)butyricacid methyl ester (18.0 q) was mixed in toluene (39.7 ml) under argon.To the reaction solution was added dropwise an 1M diisobutylaluminumhydride/toluene solution (59.1 mL) at −78° C., and the reaction solutionwas stirred at −78° C. for 2 hours. Then, the reaction solution wasgradually warmed to 0° C., and then stirred for 30 minutes. To thereaction solution was added methanol (39 mL) at −78° C. To the reactionsolution were added 30 w/v % aqueous L-tartaric acid solution (75 mL)and ethyl acetate (300 mL) under ice cooling, and the layers wereseparated. The resulting aqueous layer was extracted with ethyl acetate(100 mL). Combined organic layers were washed with 30 w/v % aqueousL-tartaric acid solution (80 mL, twice), water (80 ml, twice), andbrine, and then dried over sodium sulfate. Sodium sulfate was removedthrough a filter, and then the filtrate was concentrated under reducedpressure. The resulting residue was azeotroped with toluene to give acrude product of the title compound (18.5 g).

¹H-NMR (400 MHz, CDCl₃) 0.70 (d, J=6.94 Hz, 1.8H), 0.76 (d, J=6.94 Hz,1.8H), 0.79 (d, J=6.94 Hz, 1.2H), 0.83 (d, J=6.94 Hz, 1.2H), 1.15 (s,5.4H), 1.30 (s, 3.6H), 1.56-1.59 (m, 0.4H), 1.71-1.82 (m, 1H), 1.95 (s,1.8H), 1.98 (s, 1.2H), 2.04-2.08 (m, 0.6H), 3.86-4.13 (m, 2H), 5.13 (s,0.6H), 5.99 (s, 0.4H), 7.16-7.21 (m, 1H), 7.54-7.58 (m, 2H)

Step 4 (R)-3-Amino-3-(4-bromo-3-chloro-phenyl)-2-isopropyl-butan-1-ol

(S)-2-Methyl-propane-2-sulfinic acid[(R)-1-(4-bromo-3-chloro-phenyl)-2-hydroxymethyl-1,3-dimethyl-butyl}amide(18.5 g) was mixed in methanol (79.5 mL). To the reaction solution wasadded dropwise a 2M hydrogen chloride/methanol solution (39.8 mL) underice cooling, and the reaction solution was stirred for 1.5 hours. Thereaction solution was concentrated under reduced pressure, and to theresidue was added 10 w/v % aqueous sodium carbonate solution (50.0 mL).The reaction solution was extracted with ethyl acetate (80 mL). Theresulting aqueous layer was extracted with ethyl acetate (twice).Combined organic layers were washed with a 10 w/v % aqueous sodiumcarbonate solution (50 mL), water (50 mL), and brine, and then driedover sodium sulfate. Sodium sulfate was removed through a filter, andthen the resulting organic layer that was concentrated under reducedpressure was dried over magnesium sulfate. Magnesium sulfate was removedthrough a filter, and then the filtrate was concentrated under reducedpressure. The resulting residue was azeotroped with toluene to give acrude product of the title compound (15.7 g).

¹H-NMR (400 MHz, CDCl₃) 0.69 (d, J=6.94 Hz, 1.2H), 0.83 (d, J=6.94 Hz,1.8H), 0.85 (d, J=6.94 Hz, 1.2H), 0.87 (d, J=6.94 Hz, 1.8H), 1.36-1.47(m, 0.6H), 1.59 (s, 1.2H), 1.60 (s, 1.8H), 1.64 (ddd, J=5.32, 3.21, 2.31Hz, 0.4H), 1.66-1.77 (m, 0.4H), 1.85 (ddd, J=8.79, 3.47, 2.31 Hz, 0.6H),3.69 (dd, J=11.33, 3.47 Hz, 0.6H), 3.81 (dd, 3=11.79, 5.32 Hz, 0.4H),3.91 (dd, J=11.33, 8.79 Hz, 0.6H), 3.96 (dd, 3=11.79, 3.47 Hz, 0.4H),7.18-7.20 (m, 1H), 7.50 (d, J=2.31 Hz, 0.6H), 7.53 (d, J=2.31 Hz, 0.4H),7.58 (d, J=6.70 Hz, 0.6H), 7.60 (d, J=6.70 Hz, 0.4H.)

Step 53-{3-[(R)-1-(4-Bromo-3-chloro-phenyl)-2-hydroxymethyl-1,3-dimethyl-butyl]ureido}bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester

3-(Methoxycarbonyl) bicyclo[1.1.1]pentane-1-carboxylic acid (4.00 g) andtoluene (47.0 ml) were mixed under nitrogen, and thereto were addeddiphenylphosphoryl azide (5.58 ml) and triethylamine (3.60 mL) at roomtemperature. The reaction solution was stirred at 120° C. for 50minutes. The reaction solution was slowly added dropwise to a solutionof (R)-3-amino-3-(4-bromo-3-chloro-phenyl)-2-isopropyl-butan-1-ol (9.77g) in tetrahydrofuran (47.0 ml) over 15 minutes under ice cooling. Thereaction solution was stirred at room temperature for 1 hour. To thereaction solution was added 10 w/v % aqueous citric acid solution (100mL), and the layers were separated. The resulting organic layer waswashed with water (60 mL). Combined aqueous layers were extracted withethyl acetate (twice). Combined organic lavers were washed with 10 w/v %aqueous citric acid solution (60 MI), water (60 mL), 10 w/is aqueoussodium carbonate solution (60 mL), water (60 mL), and brine, and thendried over sodium sulfate. Sodium sulfate was removed through a filter,and then the filtrate was concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(ethyl acetate:n-hexane=7:13→100:0) to give the title compound (8.34 g).

¹H-NMR (400 MHz, CDCl₃) 0.28 (d, J=(0.94 Hz, 1.2H), 0.79 (d, J=6.94 Hz,1.8H), 0.85 (d, J=6.94 Hz, 1.8H), 0.92 (d, J=6.94 Hz, 1.2H), 1.52-1.59(m, 0.6H), 1.79-1.88 (m, 0.4H), 1.84 (s, 1.8H), 1.90 (5, 1.2H),2.12-2.18 (m, 0.4H), 2.21-2.32 (m, 0.6H), 2.29 (s, 3.6H), 2.29 (s,2.4H), 3.67 (s, 3H), 3.82-3.75 (m, 1.4H), 3.89-3.94 (m, 0.6H), 4.61 (s,0.6H), 4.70 (s, 6.92 (s, 0.4H), 7.04 (s, 0.6H), 7.13-7.16 (m, 1H), 7.44(d, J=2.31 Hz, 0.4H), 7.45 (d, J=2.08 Hz, 0.6H), 7.53 (d, J=8.55 Hz, 1H)

Step 6 3-[(S)-4-(4-Bromo-3-chloro-phenyl)-5-isopropyl-4-methyl2-oxo-3,4-dihydro-2H-pyrimidin-1-yl]-bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester

3-{3-[(R)-1-(4-Bromo-3-chloro-phenyl)-2-hydroxymethyl1,3-dimethyl-butyl]ureido}bicyclo[1.1.1]pentane-1-carboxylic acid methylester (10.0 g) and chloroform (68.4 mL) were mixed under nitrogen, andthereto were added (diacetoxyiodo)benzene (6.06 g) and2,2,6,6-tetramethylpiperidin-1-oxyl radical (0.267 g) at roomtemperature. The reaction solution was stirred at room temperature for3.5 hours, and then thereto was added 10 w/w % aqueous sodium sulfitesolution (50 mL) at room temperature. The layers were separated. Theresult nq aqueous layer was extracted with chloroform (twice). Combinedorganic layers were washed with water (50 mL) and brine, and then driedover sodium sulfate. Sodium sulfate was removed through a filter, andthen the filtrate was concentrated under reduced pressure. The resultingresidue was azeotroped with toluene. The resulting residue was mixedwith toluene (213 mL), and then thereto was added pentafluoroanilinetrifluoromethanesulfonate (0.307 g) at room temperature. The reactionsolution was stirred under warming at 100° C. for 2 hours, and thenthereto were added sodium sulfite (880 mg) and 10 w/v % aqueous sodiumcarbonate solution (50 mL) at room temperature. The layers wereseparated. The resulting aqueous layer was extracted with ethyl acetate(twice). Combined organic layers were washed with water (50 mL) andbrine, and then dried over magnesium sulfate. Magnesium sulfate wasremoved through a filter, and then the filtrate was concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (ethyl acetate:n-hexane=1:4→3:2). The resultingresidue was solidified with n-hexane, and the precipitated solid wasfiltered to give the title compound (6.35 g).

¹H-NMR (400 MHz, CDCl₃) 0.72 (d, J=6.94 Hz, 3H), 1.06 (d, 3=6.94 Hz,3H), 1.70 (s, 3H), 1.81-1.92 (m, 1H), 2.47 (s, 6H), 3.71 (s, 3H), 4.69(s, 1H), 5.85 (s, 1H), 7.20 (dd, J=8.32, 2.31 Hz, 1H), 7.51 (d, J=2.31Hz, 1H), 7.58 (d, J=8.32 Hz, 1H)

Step 73-{(S)-4-[4-(2-tert-Butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester

3-[(S)-4-(4-Bromo-3-chloro-phenyl)-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl]-bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester (5.23 g), potassiumtrans-2-tert-butyl-cyclopropyl-trifluoroborate (2.79 g)[1,1′-bis(di-tert-butyl-phosphino)ferrocene]palladium (II) dichloride(0.729 g), cesium carbonate (10.9 g), toluene (112 mL), and water (11.2mL) were mixed under argon, and the mixture was stirred at 100° C. for 2hours. To the reaction solution was added water (50 Pal) at roomtemperature, and the layers were separated. The resulting aqueous layerwas extracted with ethyl acetate (twice). Combined organic layers werewashed with water (50 ml, twice) and brine, and then dried overmagnesium sulfate. Then, thereto was added activated carbon, and themixture was stirred at room temperature for hour. Magnesium sulfate andactivated carbon were removed through a filter, and then the filtratewas concentrated under reduced pressure. The resulting residue waspurified by silica gel column chromatography. (ethylacetate:n-hexane=1:9→1:1). The resulting residue was mixed withmethanol, and thereto was added activated carbon at room temperature.The activated carbon was removed through a filter, and then the filtratewas concentrated under reduced pressure. The resulting residue waspurified by silica gel column chromatography (ethylacetate:n-hexane=1:4→1:1) to give a diastereomer mixture of the titlecompound (3.70 g). Purification with a chiral preparative column gavethe title compound (0.344 q).

Purification conditions for the preparative column are shown as follows.

Preparative apparatus: Recycling Preparative Liquid ChromatographLC-92XX NEXT SERIES, Japan Analytical Industry Co., Ltd.Column: Daicel CHIRALPAK IA 2.0 cmφ×25 cmLMobile phase: n-hexane:2-propanol=93:7Flow rate: 10.0 mL/min

Detection: UV (220 nm)

Measurement with a chiral column showed 8.3 minutes of the retentiontime for the resulting title compound (7.6 minutes of the retention timefor diastereomers of the title compound) with >99% de purity. Analyticalconditions for the chiral column are shown as follows.

Measuring apparatus: HPLC system, Shimadzu Corporation, High-PerformanceLiquid Chromatograph ProminenceColumn: Daicel CHIRALPAK IA-3 0.46 cmφ×15 cmLColumn temperature: 40° C.Mobile phase: n-hexane:2-propanol=93:7Flow rate: 1.0 mL/min

Detection: UV (220 nm)

¹H-NMR (400 MHz, CDCl₃) 0.71 (d, J=7.09 Hz, 3H), 0.77-0.83 (m, 1H),0.90-0.95 (m, 2H), 0.94 (s, 9H), 1.04 (d, J=7.09 Hz, 3H), 1.68 (s, 3H),1.85-1.92 (m, 1H), 2.07-2.12 (m, 1H), 2.48 (s, 6H), 3.71 (s, 3H), 4.56(s, 1H), 5.83 (s, 1H), 6.86 (d, J=8.31 Hz, 1H), 7.22 (dd, J=8.31, 1.96Hz, 1H), 7.37 (d, J=1.96 Hz, 1H)

Step 83-{(S)-4-[4-((1R,2R)-2-tert-Butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid

3-((S)-4-[4-(2-tert-Butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl)bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester (49.1 g), tetrahydrofuran (246 mL), and methanol (246mL) were mixed under nitrogen, and thereto was added 2N aqueous sodiumhydroxide solution (101 mL) at room temperature. The reaction solutionwas stirred at room temperature for 24 hours 30 minutes. The reactionsolution was concentrated under reduced pressure, and to the residuewere added water (400 mL) and 1N hydrochloric acid (250 mL). Then,thereto was added ethyl acetate (900 mL), and the layers were separated.The resulting aqueous layer was extracted with ethyl acetate (100 mL×2),and the organic layer was washed with water (250 mL) and brine, and thendried over sodium sulfate. Sodium sulfate was removed through a filter,and then the filtrate was concentrated under reduced pressure.

The resulting residue and acetonitrile (1000 mL) were mixed, and themixture was stirred under warming at 85° C. for 1 hour 15 minutes. Then,the mixture was stirred at room temperature for 13 hours 15 minutes, andthen the resulting solid was filtered to give a crystal of the titlecompound (Crystalline Form A; 44.7 g).

A crystal of the title compound (29.7 mg) obtained in a similar manner,ethyl acetate (297 μL), and methanol (891 μL) were mixed. Solids werecompletely dissolved, and then solvents were concentrated under reducedpressure with a rotary evaporator, affording another crystal of thetitle compound (Crystalline Form B).

The above two crystals (10.0 mg each) obtained in a similar manner andisobutyl acetate (80 μL) were mixed, and the resulting slurry wasstirred at room temperature for one week, and then the resulting solidwas filtered to give another crystal of the title compound (CrystallineForm C), which was used for a seed crystal in the following procedures.

A crude crystal (2.00 g) obtained in a similar reaction to the above andisobutyl acetate (10 mL) were mixed and stirred at 90° C. After thecrude crystal was completely dissolved, the seed crystal was added tothe mixture at 47° C. of the internal temperature, and the mixture wasstirred for 1 hour. At around 45° C. of the internal temperature,n-heptane (30 mL) was added to the reaction solution, and the mixturewas stirred for 2 hours. The mixture was stirred at room temperature for24 hours. The precipitated solid was filtered to give the title compound(Crystalline Form C; 1.7 g).

¹H-NMR (400 MHz, DMSO-Dr) 0.71 (d, J=6.94 Hz, 3H), 0.86-0.94 (m, 3H),0.90 (s, 9H), 1.04 (d, J=6.94 Hz, 3H), 1.60 (s, 3H), 1.93-2.04 (m, 2H),2.29 (s, 6H), 5.97 (s, 1H), 6.99 (d, J=7.86 Hz, 1H), 7.06 (s, 1H), 7.21(dd, J=7.86, 1.85 Hz, 1H), 7.32 (d, J=1.85 Hz, 1H), 12.44 (s, 1H)

Absolute configurations of the asymmetric carbons of the title compoundwere determined by single-crystal X-ray structural analysis.

Synthesis of Intermediate 1 Potassiumtrans-2-tert-butyl-cyclopropyl-trifluoroborate

2-(trans-2-tert-Butyl-cyclopropyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborolane(8.03 g), methanol (200 mL), and water (40 mL) were mixed undernitrogen, and thereto was added potassium hydrogen fluoride (14 g) atroom temperature. The reaction solution was stirred at 100° C. for 7hours. Then, the solution was let stand at room temperature overnight,and then concentrated under reduced pressure. The resulting residue wasazeotroped with toluene. The resulting residue and acetonitrile (120 mL)were mixed and stirred at 60° C. for 2 hours. The resulting solid wasremoved through a filter, and then the filtrate was concentrated underreduced pressure. The resulting residue and diethyl ether (80 mL) weremixed and stirred at room temperature. The precipitated solid wasfiltered to give the title compound (2.86 g).

¹H-NMR (400 MHz, DMSO-D₆) −0.94-0.87 (m, 1H), −0.26-0.23 (m, 2H),0.25-0.30 (m, 1H), 0.73 (s, 9H)

Example 1A Alternative Synthesis of3-((S)-4-[4-((1R,2R)-2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl)bicyclo[1.1.1]pentane-1-carboxylicacid Step 1(4S,5S)-2-((E)-3,3-Dimethyl-1-butenyl)-[1,3,2]dioxaborolane-4,5-dicarboxylicacid bis-dimethylamide

2M Dibromoborane-dimethyl sulfide complex/toluene solution (1250 mL) wasmixed in toluene (1169 mL). To the reaction solution was added dropwise3,3-dimethyl-1-butyne (338 mL) under water cooling for 45 minutes. Thereaction solution was stirred under water cooling for 2.5 hours. Thereaction solution was cooled by addition of ice into a water bathagainst delayed exotherm. The reaction solution was added dropwise to amixed solution of 8N aqueous solution of potassium hydroxide (969 mL)and water (2922 mL) under ice cooling for 30 minutes. Toluene (1461 mL)and tetrahydrofuran (292 mL) were added to the aqueous layer, andthereto was added dropwise 6N hydrochloric acid (417 mL) under icecooling for 20 minutes. Thereto was added sodium chloride (584 g), andthe organic layer was separated.

The organic layer was mixed with toluene (1581 mL), and thereto wasadded L-tartaric acid-N,N,N′,N′-tetramethylamide (562 g). The reactionsolution was stirred for 2 hours under heating at 130° C. The organiclayer was separated and concentrated under reduced pressure. Solidsprecipitated during the concentration were removed with a filter, andthe filtrate was concentrated under reduced pressure to give a crudeproduct of the title compound (599 g).

¹H NMR (400 MHz, DMSO-D₆) 1.00 (s, 9H), 2.87 (s, 6H), 3.06 (s, 6H), 5.30(d, J=18.24 Hz, 1H), 5.44 (s, 2H), 6.64 (d, J=18.24 Hz, 1H)

Step 2 (1R,2R)-2-tert-Butylcyclopropylboronic acid

1.1M Diethylzinc/n-hexane solution (1892 mL) was mixed in methyltert-butyl ether (838 mL) under argon. To the reaction solution wasadded dropwise chloroiodomethane (301 mL) under being cooled with dryice-acetone for 35 minutes. The reaction solution was stirred underbeing cooled with dry ice-acetone for 30 minutes. Then, thereto wasadded L-tartaric acid-N,N,N′,N′-tetramethylamide (70.8 g) under beingcooled with dry ice-acetone, and the mixture was stirred for 20 minutes.To the reaction solution was added dropwise a mixed solution of(4S,5S)-2-((E)-3,3-dimethyl-1-butenyl)-[1,3,2]dioxaborolane-4,5-dicarboxylicacid bis-dimethylamide (222 g) in methyl tert-butyl ether (838 mL) underbeing cooled with dry ice-acetone for 30 minutes. The reaction solutionwas stirred under being cooled with dry ice-acetonitrile for 4.5 hours.Then, thereto was added a 30 w/v % aqueous solution of citric acid (1438mL) under being cooled with dry ice-acetonitrile over 10 minutes. To thereaction solution were added ethyl acetate (1047 mL) and sodium chloride(335 g), and the organic layer was washed with water (1047 mL) and a 25w/v % aqueous solution of sodium chloride (1047 mL) and then dried overmagnesium sulfate. Magnesium sulfate was removed with a filter, and thenthe filtrate was concentrated under reduced pressure to give a crudeproduct (82.6 g).

The resulting crude product (82.6 g) was mixed with an 8N aqueoussolution of potassium hydroxide (87 mL) and water (150 mL). The reactionsolution was added dropwise to 6N hydrochloric acid (117 mL) under icecooling for 1 hour. The reaction solution was stirred under ice coolingfor 1 hour. The precipitated solid was filtered to give the titlecompound (68.8 g).

¹H NMR (400 MHz, DMSO-D₆)-0.46 (td, J=5.98, 9.27 Hz, 1H), 0.30-0.37 (m,2H), 0.70-0.82 (m, 111), 0.79 (s, 9H), 12.94 (br s, 2H)

Step 3 Methyl 4-((1R,2R)-2-tert-butylcyclopropyl)-3-chlorobenzoate

Potassium carbonate (1329 g) was mixed in water (1600 mL) under argon.To the reaction were added toluene (4800 mL), methyl4-bromo-3-chlorobenzoate (800 g), (1R,2R)-2-tert-butylcyclopropylboronicacid (1454 g), and bis(triphenylphosphine)palladium (II) dichloride (112g), and the reaction solution was stirred for 4.5 hours under heating at80° C. The reaction solution was cooled to room temperature. To thereaction solution was added 0.5N aqueous solution of potassium hydroxide(3200 mL) under heating at 60° C., and the mixture was stirred for 2hours. The organic layer was separated, and then the aqueous layer wasextracted with toluene (1200 mL). Combined organic layers were washedwith water (3200 mL), and then thereto was added Celite (800 g). Themixture was stirred at room temperature for 1 hour. Celite was removedwith a filter, and then the filtrate was concentrated under reducedpressure. To the resulting residue was added isopropanol (2000 mL), andthe mixture was concentrated under reduced pressure to give a crudeproduct of the title compound (1061 g).

¹H NMR (400 MHz, DMSO-D₆) 0.91 (s, 9H), 1.02-1.07 (m, 3H), 2.13 (td,J=8.07, 5.68 Hz, 1H), 3.84 (s, 3H), 7.15 (d, J=8.37 Hz, 1H), 7.79 (dd,J=8.37, 1.79 Hz, 1H), 7.89 (d, J=1.79 Hz, 1H)

Step 4 4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid

Methyl 4-((1R,2R)-2-tert-butylcyclopropyl)-3-chlorobenzoate (1061 g) wasmixed in isopropanol (4600 mL). To the reaction solution was added 2Naqueous solution of sodium hydroxide (4000 mL), and the reactionsolution was stirred at room temperature overnight. To the reactionsolution was added activated carbon (160 g), and the mixture was stirredfor 30 minutes. After filtration with Celite, 2N hydrochloric acid (4100mL) was added to the filtrate under ice cooling. The filtrate wasextracted with ethyl acetate (4800 mL). The aqueous layer was extractedwith ethyl acetate (1200 mL), and combined organic layers were washedwith saturated aqueous solution of sodium chloride (1600 mL) and thendried over magnesium sulfate. Magnesium sulfate was removed with afilter, and then the filtrate was concentrated under reduced pressure.To the resulting residue was added acetonitrile (1600 mL), and themixture was concentrated under reduced pressure to give a crude productof the title compound (920 g).

¹H NMR (400 MHz, DMSO-D₆) 0.92 (s, 9H), 1.00-1.06 (m, 3H), 2.10-2.15 (m,1H), 7.12 (d, J=7.77 Hz, 1H), 7.77 (d, J=7.77 Hz, 1H), 7.87 (s, 1H),13.11 (br s, 1H)

Step 5 4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid

4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid (920 g) wasmixed in acetonitrile (7200 mL) and water (4000 mL). The reactionsolution was stirred under heating at an internal temperature of 67° C.,and 4-((1R,2R)-2-tert-butylcyclopropyl)-3-chlorobenzoic acid wasconfirmed to be completely dissolved. Then, water (3700 mL) was added tothe reaction solution, and thereto was added a seed crystal. The mixturewas stirred for 6 hours under heating at 60° C. and then stirredovernight with being cooled slowly to room temperature. The precipitatedsolid was filtered to give the title compound (518 g). In this step, theseed crystal was used to obtain the crystal efficiently, but the crystalcan be obtained without the seed crystal in similar procedures to thisstep.

¹H NMR (400 MHz, DMSO-D₆) 0.92 (s, 9H), 1.00-1.06 (m, 3H), 2.10-2.15 (m,1H), 7.12 (d, J=7.77 Hz, 1H), 7.77 (d, J=7.77 Hz, 1H), 7.87 (s, 1H),13.11 (br s, 1H)

Step 6 4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid(S)-phenethylamine salt

4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid (790 g) wasmixed in isopropyl acetate (7900 mL). To the mixture was added dropwise(S)-(−)-phenethylamine (417 g) under heating at an internal temperatureof 87° C. for 10 minutes. Then, a seed crystal was added thereto. Themixture was stirred for 2 hours under heating at 83° C. and then stirredovernight with being cooled slowly to room temperature. The precipitatedsolid was filtered to give the title compound (1087 g). In this step,the seed crystal was used to obtain the crystal efficiently, but thecrystal can be obtained without the seed crystal in similar proceduresto this step.

¹H NMR (400 MHz, DMSO-D₆) 0.91 (s, 9H), 0.94-1.00 (m, 3H), 1.41 (d,J=6.58 Hz, 3H), 2.05-2.09 (m, 1H), 4.25 (q, J=6.58 Hz, 1H), 6.98 (d,J=8.07 Hz, 1H), 7.27-7.30 (m, 1H), 7.35-7.38 (m, 2H), 7.45-7.47 (m, 2H),7.70 (d, J=8.07 Hz, 1H), 7.81 (s, 1H)

Step 7 4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid(S)-phenethylamine salt

4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid(S)-phenethylamine salt (1087 g) was mixed in isopropyl acetate (7600mL). To the reaction solution was added (S)-(−)-phenethylamine (35.2 g).The mixture was stirred for 2 hours under heating at 83° C. and thenstirred overnight with being cooled slowly to room temperature. Theprecipitated solid was filtered to give the title compound (1044 g).

¹H NMR (400 MHz, DMSO-D₆) 0.91 (s, 9H), 0.94-1.00 (m, 3H), 1.41 (d,J=6.58 Hz, 3H), 2.05-2.09 (m, 18), 4.25 (q, J=6.58 Hz, 1H), 6.98 (d,J=8.07 Hz, 1H), 7.27-7.30 (m, 1H), 7.35-7.38 (m, 2H), 7.45-7.47 (m, 2H),7.70 (d, J=8.07 Hz, 1H), 7.81 (s, 1H)

Step 8 4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid

4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid(S)-phenethylamine salt (1044 g) was mixed in acetonitrile (5200 mL) andwater (3800 mL). Thereto was added 2N hydrochloric acid (1470 mL), andthe mixture was stirred for 2 hours under heating at 80° C. and thenstirred overnight with being cooled slowly to room temperature. Then,thereto was added dropwise water (4200 mL) for 1 hour, and then themixture was stirred at room temperature for 2 hours. The precipitatedsolid was filtered to give the title compound (697 g).

¹H NMR (400 MHz, DMSO-D_(E)) 0.92 (s, 9H), 1.00-1.06 (m, 3H), 2.10-2.15(m, 1H), 7.12 (d, J=7.77 Hz, 1H), 7.77 (d, J=7.77 Hz, 1H), 7.87 (s, 1H),13.11 (br s, 1H)

Step 94-((1R,2R)-2-tert-Butylcyclopropyl)-3-chloro-N-methoxy-N-methylbenzamide

4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chlorobenzoic acid (1000 g) wasmixed in N-methylpyrrolidone (5000 mL). To the reaction solution wereadded N,O-dimethylhydroxylamine hydrochloride (540 g),1-hydroxybenzotriazole monohydrate (60.6 g), and sodium hydrogencarbonate (465 g). To the mixture was addedN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1099 g)under ice cooling, and the mixture was stirred for 1 hour. The mixturewas stirred at room temperature for 1 hour, and then thereto were addedcyclopentyl methyl ether (5000 mL) and water (2500 mL). The aqueouslayer was extracted with cyclopentyl methyl ether (400 mL), and combinedorganic layers were washed with water (2500 mL) and 20 w/v % aqueoussolution of sodium chloride (2500 mL) and then dried over sodiumsulfate. Sodium sulfate was removed with a filter, and then concentratedunder reduced pressure. To the resulting residue was added cyclopentylmethyl ether (3000 mL), and the mixture was concentrated under reducedpressure to give a crude product of the title compound (1356 g).

¹H NMR (400 MHz, DMSO-D₆) 0.92 (s, 9H), 0.95-1.04 (m, 3H), 2.07-2.12 (m,1H), 3.25 (s, 3H), 3.55 (s, 3H), 7.08 (d, J=7.77 Hz, 1H), 7.48 (d,J=7.77 Hz, 1H), 7.61 (s, 1H)

Step 10 1-[4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chloro-phenyl]ethanone

4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chloro-N-methoxy-N-methylbenzamide(1356 g) was mixed in tetrahydrofuran (5900 mL). To the reactionsolution was added dropwise 3M methylmagnesium chloride/tetrahydrofuransolution (1700 mL) under ice cooling for 2 hours, and then the mixturewas stirred for 1 hour. To the reaction solution was added isopropanol(117 mL), and then thereto was added dropwise 2N hydrochloric acid (5900mL). Toluene (6000 mL) was added thereto, and the organic layer waswashed with 20 w/v % aqueous solution of sodium chloride (6000 mL) andwater (3600 mL). Then, the solution was dried over sodium sulfate.Sodium sulfate was removed with a filter, and then concentrated underreduced pressure. To the resulting residue was added cyclopentyl methylether (3500 mL), and the mixture was concentrated under reduced pressureto give a crude product of the title compound (1193 g).

¹H NMR (400 MHz, DMSO-D) 0.92 (s, 9H), 1.02-1.07 (m, 3H), 2.11-2.16 (m,1H), 2.55 (s, 3H), 7.14 (d, J=8.07 Hz, 1H), 7.79 (d, J=8.07 Hz, 1H),7.92 (s, 1H)

Step 11 (S)-2-Methyl-propane-2-sulfinic acid{1-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]eth-(E)-yliden}amide

1-[4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chloro-phenyl]ethanone (207 g)and (S)-(−)-2-methyl-propane-2-sulfinic acid amide (116 g) were mixed incyclopentyl methyl ether (1000 mL). To the reaction solution was addedtetraethyl orthotitanate (335 mL), and the reaction solution was stirredfor 3 hours under heating at 110° C. To the reaction solution was addedmethanol (1000 mL) under water cooling, and the mixture was stirred for10 minutes. Thereto was added dropwise a mixed aqueous solution of 28w/v % aqueous solution of ammonia (100 mL), L-lactic acid (178 mL), andwater (1000 mL) at an internal temperature of 30° C. or below for 15minutes. The mixture was stirred at room temperature for 4 hours, andthen let stand overnight. To the aqueous layer was added cyclopentylmethyl ether (1000 mL), and combined organic layers were washed with 25w/v % aqueous solution of sodium chloride (1000 mL) and then dried oversodium sulfate. Sodium sulfate was removed with a filter, and then thefiltrate was concentrated under reduced pressure. To the residue wasadded n-hexane (400 mL), and the mixture was concentrated under reducedpressure. To the residue was added n-hexane (400 mL) additionally, andthen the mixture was concentrated under reduced pressure. Then, theretowas added dimethyl sulfoxide (500 mL), and the mixture was concentratedunder reduced pressure. To the residue was added dimethyl sulfoxide (500mL), and to the mixture was added dropwise water (50 mL) at roomtemperature. The mixture was stirred at room temperature for 30 minutes,and then thereto was added dropwise water (150 mL). The mixture wasstirred at room temperature for 12 hours, and then the precipitatedsolid was filtered to give the title compound (261 g).

¹H NMR (400 MHz, DMSO-DG) 0.91 (s, 9H), 1.01-1.05 (m, 3H), 1.21 (s, 9H),2.12 (td, J=7.63, 5.78 Hz, 1H), 2.69 (s, 3H), 7.11 (d, J=8.55 Hz, 1H),7.75 (dd, J=8.55, 1.62 Hz, 1H), 7.87 (d, J=1.62 Hz, 1H)

Step 12 Methyl(R)-3-[4-((1R,2R)-2-tert-Butylcyclopropyl)-3-chloro-phenyl]-2-isopropyl-3-((S)-2-methyl-propane-2-sulfinylamino)butanoate

2M Lithium diisopropylamide/tetrahydrofuran/n-heptane/ethylbenzenesolution (740 mL) was mixed in tetrahydrofuran (786 mL) under nitrogenflow. To the reaction solution was added dropwise a mixed solution ofmethyl 3-methyl-butanoate (224 mL) in tetrahydrofuran (524 mL) undercooling at −78° C. for 45 minutes, and the reaction solution was stirredunder cooling at −78° C. for 2 hours 30 minutes. To the reactionsolution was added dropwise a mixed solution of(S)-2-methyl-propane-2-sulfinic acid{1-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]eth-(E)-yliden}amide(261 g) in tetrahydrofuran (393 mL) for 40 minutes, and the reactionsolution was stirred for 4 hours. To the reaction solution were addeddropwise sequentially methanol (131 mL) and water (131 mL) within 5minutes, and the mixture was stirred at room temperature for 14 hours.To the reaction solution was added water (786 ml) under water cooling,and the mixture was separated. The aqueous layer was extracted withtoluene (1048 mL). Combined organic layers were washed sequentially with1N sulfuric acid (1309 mL), water (786 mL), 5 w/v % aqueous solution ofsodium hydrogen carbonate (786 mL), and 25 w/v % aqueous solution ofsodium chloride (786 mL) and then dried over sodium sulfate. Sodiumsulfate was removed with a filter, and then concentrated under reducedpressure. To the resulting residue was added toluene (262 mL), and themixture was concentrated under reduced pressure to give a crude productof the title compound (348 g).

¹H NMR (400 MHz, DMSO-D₆) 0.76-0.92 (m, 18H), 1.15 (s, 6.3H), 1.20 (s,2.7H), 1.75 (s, 0.9H), 1.76 (s, 2.1H), 1.93-2.04 (m, 2H), 2.68 (d,J=3.59 Hz, 0.3H), 2.93 (d, J=3.89 Hz, 0.7H), 3.52 (s, 2.1H), 3.58 (s,0.9H), 5.15 (s, 0.7H), 5.44 (s, 0.3H), 6.97 (d, J=8.37 Hz, 0.71H), 6.98(d, J=8.37 Hz, 0.3H), 7.31 (d, J=8.37 Hz, 1H), 7.46 (s, 0.7H), 7.49 (s,0.3H)

Step 13 (S)-2-Methyl-propane-2-sulfinic acid[(R)-1-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]-2-hydroxymethyl-1,3-dimethyl-butyl}amide

To 1M diisobutylaluminum hydride/toluene solution (2932 mL) was addeddropwise a mixed solution of methyl(R)-3-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]-2-isopropyl-3-((S)-2-methyl-propane-2-sulfinylamino)butanoate(348 g) in toluene (1392 mL) under cooling at −78° C. for 1 hour undernitrogen flow. The reaction solution was stirred under cooling at −78°C. for 2 hours. The reaction solution was added to a mixed solution ofcitric acid monohydrate (622 g) in water (2088 mL) at an internaltemperature of 20° C. or below under ice cooling. The mixture wasstirred at room temperature for 15 hours, and then separated. Theaqueous layer was extracted with toluene (1044 mL). Combined organiclayers were washed sequentially with water (1740 mL) and 25 w/v %aqueous solution of sodium chloride (1740 mL), and then dried oversodium sulfate. Sodium sulfate was removed with a filter, and then thefiltrate was concentrated under reduced pressure to give a crude productof the title compound (645 g).

¹H NMR (400 MHz, DMSO-D₆) 0.65 (d, J=7.09 Hz, 2.1H), 0.68-0.71 (m, 3H),0.75 (d, J=7.09 Hz, 0.9H), 0.82-0.95 (m, 12H), 1.05 (s, 6.3H), 1.18 (s,2.7H), 1.73 (s, 2.1H), 1.79 (s, 0.9H), 1.84-1.88 (m, 0.7H), 1.99-2.05(m, 1.3H), 3.65-3.80 (m, 2H), 5.14 (t, J=3.55 Hz, 0.3H), 5.50 (t, J=3.91Hz, 0.7H), 6.03 (s, 0.3H), 6.56 (s, 0.7H), 6.93-6.99 (m, 1H), 7.28-7.34(m, 1H), 7.38 (d, J=1.96 Hz, 0.7H), 7.43 (d, J=1.71 Hz, 0.311)

Step 14(R)-3-Amino-3-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]-2-isopropyl-butan-1-olhydrochloride

(S)-2-Methyl-propane-2-sulfinic acid[(R)-1-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]-2-hydroxymethyl-1,3-dimethyl-butyl)amide(327 g) was mixed in toluene (1309 mL). To the reaction solution wasadded dropwise 2M hydrogen chloride/methanol solution (740 mL) for 20minutes under ice cooling, and the mixture was stirred for 2 hours underwater cooling. The reaction solution was concentrated under reducedpressure, and to the residue was added diisopropyl ether (250 mL). Themixture was concentrated under reduced pressure, and then to the residuewas added diisopropyl ether (4155 mL). The mixture was stirred for 6hours under heating to 90° C. The reaction solution was stirred for 8hours with being cooled slowly to room temperature, and then theprecipitated solid was filtered to give the title compound (195 g).

¹H NMR (400 MHz, DMSO-D_(E)) 0.64 (d, J=6.88 Hz, 0.9H), 0.67 (d, J=6.88Hz, 2.1H), 0.78 (d, J=6.88 Hz, 2.1H), 0.91 (s, 9H), 0.94-0.98 (m, 3H),1.03 (d, J=6.88 Hz, 0.9H), 1.33-1.40 (m, 0.7H), 1.66 (s, 0.9H), 1.68 (s,2.1H), 1.92-2.08 (m, 2.3H), 3.31-3.35 (m, 0.3H), 3.50-3.55 (m, 0.3H),3.56-3.77 (m, 1.4H), 5.14 (br s, 0.3H), 5.29 (br s, 0.7H), 7.06 (d,J=8.37 Hz, 0.3H), 7.08 (d, J=8.37 Hz, 0.7H), 7.43 (d, J=8.37 Hz, 0.3H),7.49 (d, J=8.37 Hz, 0.7H), 7.60 (s, 1H), 8.31 (br s, 2.1H), 8.49 (br s,0.9H)

Step 15 Methyl3-{3-[(R)-1-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]-2-hydroxymethyl-1,3-dimethyl-butyl]ureido}bicyclo[1.1.1]pentane-1-carboxylate

3-(Methoxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid (63.8 g) wasmixed in toluene (761 mL) under argon flow, and thereto was addeddropwise triethylamine (57.5 mL) over 5 minutes or above under watercooling. Thereto was added dropwise diphenylphosphoryl azide (89 mL)over 7 minutes or above under water cooling, and the mixture was stirredfor 30 minutes. The mixture was stirred for 1 hour 30 minutes withheating to 100° C. The reaction solution was added dropwise for 37minutes under ice cooling to a suspension which was prepared by mixing(R)-3-amino-3-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]-2-isopropyl-butan-1-olhydrochloride (117 g) with tetrahydrofuran (819 mL) followed by dropwiseaddition of triethylamine (47.9 mL) to the mixture over 5 minutes orbelow under water cooling. The mixture was stirred for 30 minutes underice cooling, and then stirred at room temperature for 1 hour 30 minutes.To the mixture was added dropwise 1N hydrochloric acid (1170 mL) over 15minutes or above under ice cooling, and then the mixture was separated.The organic layer was washed sequentially with water (585 mL), 5 w/v %aqueous solution of sodium hydrogen carbonate (585 mL, twice), water(585 mL), and 25 w/v % aqueous solution of sodium chloride (585 mL), andthen dried over sodium sulfate. Sodium sulfate was removed with afilter, and then the filtrate was concentrated under reduced pressure.To the resulting residue was added cyclopentyl methyl ether, and themixture was concentrated under reduced pressure. This procedure wasrepeated twice to give a crude product of the title compound (192 g).

¹H NMR (400 MHz, CDCl₃) 0.27 (d, J=6.94 Hz, 0.9H), 0.73-0.79 (m, 1H),0.82 (d, J=6.94 Hz, 2.1H), 0.86-0.99 (m, 2.9H), 0.89 (d, J=6.94 Hz,2.1H), 0.93 (s, 2.7H), 0.94 (s, 6.3H), 1.49-1.55 (m, 1H), 1.84 (s,2.1H), 1.89 (s, 0.9H), 2.07 (dt, J=8.79, 5.09 Hz, 1H), 2.27 (s, 4.2H),2.28 (s, 1.8H), 2.36-2.29 (m, 1H), 3.66 (s, 2.1K), 3.69 (s, 0.9H),3.73-3.82 (m, 1.3H), 3.91 (dd, J=10.98, 7.51 Hz, 0.7H), 4.52 (s, 0.7H),4.59 (s, 0.3H), 6.71 (s, 0.3H), 6.82-6.85 (m, 1H), 6.92 (s, 0.7H),7.14-7.18 (m, 1H), 7.30-7.31 (m, 1H)

Step 16 Methyl3-((S)-4-[4-((1R,2R)-2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl)bicyclo[1.1.1]pentane-1-carboxylate

(Diacetoxyiodo)benzene (111 g) and 2,2,6,6-tetramethylpiperidine-1-oxylradical (2.44 g) were mixed in acetic acid (632 mL) under nitrogen flow,and to the mixture was added dropwise a mixed solution of methyl3-{3-[(R)-1-[4-((1R,2R)-2-tert-butylcyclopropyl)-3-chloro-phenyl]-2-hydroxymethyl-1,3-dimethyl-butyl]ureido}bicyclo[1.1.1]pentane-1-carboxylate(158 g) and 2,2,6,6-tetramethylpiperidine-1-oxyl radical (2.44 g) incyclopentyl methyl ether (632 mL) at room temperature for 20 minutes.The reaction solution was stirred at room temperature for 17 hours, andthen thereto was added dropwise trifluoroacetic acid (94 mL) for 9minutes under water cooling. The reaction solution was stirred at roomtemperature for 3 hours 30 minutes, and then thereto was added 10 w/w %aqueous solution of sodium sulfite (474 mL) under water cooling. Themixture was stirred for 45 minutes. To the mixture was added n-heptane(790 mL), and the mixture was separated. The aqueous layer was extractedwith n-heptane (474 mL). Combined organic layers were washed with water(790 mL, twice) and then dried over sodium sulfate. Sodium sulfate wasremoved with a filter, and then the filtrate was concentrated underreduced pressure. To the resulting residue was added toluene, and themixture was concentrated under reduced pressure. This procedure wasrepeated twice. To the resulting residue was added isopropanol, and themixture was concentrated under reduced pressure. This procedure wasrepeated three times to give a crude product of the title compound (197g).

¹H-NMR (400 MHz, CDCl₃) 0.71 (d, J=7.09 Hz, 3H), 0.77-0.83 (m, 1H),0.90-0.95 (m, 2H), 0.94 (s, 9H), 1.04 (d, J=7.09 Hz, 3H), 1.68 (s, 3H),1.85-1.92 (m, 1H), 2.07-2.12 (m, 1H), 2.48 (s, 6H), 3.71 (s, 3H), 4.56(s, 1H), 5.83 (s, 1H), 6.86 (d, J=8.31 Hz, 1H), 7.22 (dd, J=8.31, 1.96Hz, 1H), 7.37 (d, J=1.96 Hz, 1H)

Step 173-{(S)-4-[4-((1R,2R)-2-tert-Butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid

Methyl3-((S)-4-[4-((1R,2R)-2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl)bicyclo[1.1.1]pentane-1-carboxylate(197 g) was mixed in isopropanol (437 mL) under nitrogen flow, and tothe mixture was added dropwise 2N aqueous solution of sodium hydroxide(376 mL) for 12 minutes under water cooling. The reaction solution wasstirred at room temperature for 3 hours, and then thereto was addedn-heptane (730 mL). The mixture was separated. The aqueous layer waswashed with n-heptane (730 mL). The aqueous layer was mixed in methyltert-butyl ether (730 mL), and thereto was added dropwise 2Nhydrochloric acid (584 mL) under ice cooling. Then, the mixture wasseparated. The organic layer was washed with water (438 mL, twice) andthen dried over sodium sulfate. Sodium sulfate was removed with afilter, and then the filtrate was concentrated under reduced pressure.To the resulting residue was added acetonitrile, and the mixture wasconcentrated under reduced pressure to give a crude product of the titlecompound (152 g).

The resulting crude product was mixed in acetonitrile (2127 mL) undernitrogen flow, and the mixture was stirred for 6 hours under heating at90° C. The reaction solution was stirred for 17 hours with being cooledslowly to room temperature, and then the precipitated solid was filteredto give Crystalline Form A (110 g) of the title compound.

The resulting crude crystal was mixed in isobutyl acetate (659 mL) andstirred under heating at 105° C. After the crude crystal was completelydissolved, hot filtration was performed. The filter was washed withisobutyl acetate (200 mL), and the filtrate was stirred under heating at105° C. so that the precipitated solid was completely dissolved. Thereaction solution was stirred for 3 hours with being cooled slowly underheating at 50° C., and then a seed crystal was added to the solution.The reaction solution was stirred for 1 hour 50 minutes with beingcooled slowly under heating at 45° C. To the reaction solution was addeddropwise n-heptane (2601 mL) for 56 minutes under heating at 55° C., andthen the mixture was stirred for 2 hours. The reaction solution wasstirred for 18 hours with being cooled slowly to room temperature, andthen the precipitated solid was filtered to give Crystalline Form C (153g) of the title compound.

¹H NMR (400 MHz, DMSO-D₆) 0.71 (d, J=6.94 Hz, 3H), 0.86-0.94 (m, 3H),0.90 (s, 9H), 1.04 (d, J=6.94 Hz, 3H), 1.60 (s, 3H), 1.93-2.04 (m, 2H),2.29 (s, 6H), 5.97 (s, 1H), 6.99 (d, J=7.86 Hz, 1H), 7.06 (s, 1H), 7.21(dd, J=7.86, 1.85 Hz, 1H), 7.32 (d, J=1.85 Hz, 1H), 12.44 (s, 1H)3-{(S)-4-[4-((1R,2R)-2-tert-Butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid (50.0 mg) was mixed in toluene (0.1 mL) and n-heptane (0.1 mL), andthe mixture was stirred under heating at 80° C. so that solids werecompletely dissolved. A seed crystal was added to the mixture, and thereaction solution was stirred for 6 hours with being cooled slowly toroom temperature. Then, the precipitated solid was filtered to giveCrystalline Form B (45.0 mg) of the title compound.

3-{(S)-4-[4-((1R,2R)-2-tert-Butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid (9.71 g) was mixed in acetone (87.5 mL) and water (29 mL), and themixture was stirred for 2 hours under heating at 70° C. so that solidswere completely dissolved. The reaction solution was stirred for 2 hoursunder heating at 42.5° C. with being cooled slowly. The reactionsolution was stirred for 15 hours with being cooled slowly to roomtemperature, and then the precipitated solid was filtered to giveCrystalline Form D (7.52 g) of the title compound.

A crystal of the title compound (Crystalline Form D; 50.0 mg) was mixedin acetone (0.375 mL) and water (0.125 mL), and the mixture was stirredunder heating at 58° C. until solids were completely dissolved. Thereaction solution was stirred for 8 days with being cooled to roomtemperature, and then the resulting solid was filtered to give anothercrystal (Crystalline Form E; 18.6 mg) of the title compound, which wasused for a seed crystal in the following procedure.

3-{(S)-4-[4-((1R,2R)-2-tert-Butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid (500 mg) was mixed in acetone (3.6 mL) and water (0.9 mL), and themixture was stirred under heating at 70° C. so that solids werecompletely dissolved. The seed crystal was added to the reactionsolution with heating at 50° C., and the reaction solution was stirredfor 1 hour. Thereto was added water (3 mL) with heating at 50° C., andthe mixture was stirred for 2 hours. The reaction solution was stirredfor 3 hours with being cooled slowly to room temperature, and then theprecipitated solid was filtered to give Crystalline Form E (483 mg) ofthe title compound.

Example 2 Synthesis of3-{(S)-5-((S)-sec-butyl)-4-[4-(2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[11.1.1]pentane-1-carboxylicacid Step 1(S)-2-[(R)-1-(4-Bromo-3-chloro-phenyl)-1-((S)-2-methyl-propane-2-sulfinylamino)-ethyl]-3-methylpentanoicacid ethyl ester

2M Lithium diisopropylamide/tetrahydrofuran/n-heptane/ethylbenzenesolution (45 mL) was mixed in tetrahydrofuran (45 mL) under argon. Tothe reaction solution was added dropwise a mixed solution of(S)-3-methyl-pentanoic acid ethyl ester (12.9 g) in tetrahydrofuran (30mL) under cooling at −78° C., and the reaction solution was stirred at−78° C. for 2 hours. To the reaction solution was added dropwise a mixedsolution of (S)-2-methyl-propane-2-sulfinic acid[1-(4-bromo-3-chloro-phenyl)-eth-(E)-yliden]-amide (15.0 g) intetrahydrofuran (40 mL), and the reaction solution was stirred undercooling at −78° C. for 2 hours. To the reaction solution was addedsaturated aqueous ammonium chloride solution, and the mixture wasstirred at room temperature. The layers of the reaction solution wereseparated, and the aqueous layer was extracted with ethyl acetate(twice). Combined organic layers were washed with brine, and then driedover sodium sulfate. Sodium sulfate was removed through a filter, andthen the filtrate was concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography (toluene:ethylacetate=4:1) to give a mixture of diastereomers of the title compound(8.9 g).

¹H-NMR (400 MHz, CDCl₃) 0.66 (t, J=7.17 Hz, 2.4H), 0.87 (t, J=7.17 Hz,0.6H), 0.92 (d, J=6.94 Hz, 0.6H), 1.00 (d, J=6.94 Hz, 2.4H), 1.05-1.13(m, 1.6H), 1.19 (t, J=7.17 Hz, 0.6H), 1.24 (s, 1.8H), 1.27 (t, J=7.17Hz, 2.4H), 1.32-1.35 (m, 7.6H), 1.42-1.52 (m, 0.8H), 1.67-1.73 (m,0.2H), 1.88 (s, 0.6H), 1.90 (s, 2.4H), 2.52 (d, J=3.01 Hz, 0.8H), 2.82(d, J=3.70 Hz, 0.2H), 4.07 (q, J=7.17 Hz, 0.4H), 4.18 (q, J=7.17 Hz,1.6H), 5.04 (s, 0.2H), 5.49 (s, 0.8H), 7.18-7.21 (m, 1H), 7.55-7.59 (m,2H)

Step 2 (S)-2-Methyl-propane-2-sulfinic acid[(1R,3S)-1-(4-bromo-3-chloro-phenyl)-2-hydroxymethyl-1,3-dimethyl-pentyl}amide

To an 1M diisobutylaluminum hydride/toluene solution (202 mL) was addeddropwise a mixed solution of(S)-2-[(R)-1-(4-bromo-3-chloro-phenyl)-1-((S)-2-methyl-propane-2-sulfinylamino)-ethyl]-3-methylpentanoicacid ethyl ester (24.3 g) in toluene (100 mL) under argon under coolingat −78° C. The reaction solution was stirred under cooling at −78° C.for 2 hour 25 minutes. Methanol (24.5 mL) was added to the reactionsolution under cooling at −78° C. To the reaction solution was addedsaturated aqueous (+)-potassium sodium tartrate solution (250 mL) undercooling at −78° C., and the mixture was stirred at room temperature for2 hours. The layers of the reaction solution were separated, and theaqueous layer was extracted with toluene (300 mL). Combined organiclayers were washed with saturated aqueous (+)-potassium sodium tartratesolution (200 mL), water (200 mL), and brine (200 mL), and then driedover sodium sulfate. Sodium sulfate was removed through a filter, andthen the filtrate was concentrated under reduced pressure. The resultingresidue was azeotroped with toluene to give a crude mixture ofdiastereomers of the title compound (24.7 g).

¹H-NMR (400 MHz, CDCl₃) 0.67-0.95 (m, 6H), 1.14 (s, 3.6H), 1.20-1.27 (m,2H), 1.29 (s, 5.4H), 1.35-1.50 (m, 1H), 1.80-1.82 (m, 0.6H), 1.92 (s,1.2H), 1.94 (s, 1.8H), 2.02-2.05 (m, 0.4H), 2.73-2.80 (m, 0.6H),3.18-3.26 (m, 0.4H), 3.78-4.08 (m, 2H), 5.35 (s, 0.6H), 6.23 (s, 0.4H),7.16-7.20 (m, 1H), 7.52-7.62 (m, 2H)

Step 3(S)-2-[(R)-1-Amino-1-(4-bromo-3-chloro-phenyl)-ethyl]-3-methyl-pentan-1-ol

2-Methyl-propane-2-sulfinic acid[(1R,3S)-1-(4-bromo-3-chloro-phenyl)-2-hydroxymethyl-1,3-dimethyl-pentyl)amide(24.7 g) was mixed in toluene (124 mL). To the reaction solution wasadded dropwise 2M hydrogen chloride/methanol solution (50.4 mL) underice cooling, and the reaction 1.0 solution was stirred for 3 hours undercooling with a water bath. To the reaction solution was added 4N aqueoussodium hydroxide solution (25 mL) under ice cooling. The pH value of thereaction solution was adjusted to 12 by addition of 2N aqueous sodiumhydroxide solution under ice cooling. The layers of the reactionsolution were separated, and the aqueous layer was extracted withtoluene (200 mL). Combined organic layers were washed with brine, andthen dried over sodium sulfate. Sodium sulfate was removed through afilter, and then the filtrate was concentrated under reduced pressure.The resulting residue was azeotroped with toluene to give a crudemixture of diastereomers of the title compound (19.5 g).

¹H-NMR (400 MHz, CDCl₃) 0.55 (d, J=6.94 Hz, 1.8H), 0.75 (t, J=7.28 Hz,1.2H), 0.79 (t, J=7.17 Hz, 1.8H), 0.84 (d, J=7.51 Hz, 1.2H), 0.86-1.07(m, 1H), 1.20-1.29 (m, 1H), 1.35-1.45 (m, 1H), 1.59 (s, 1.2H), 1.62 (s,1.8H), 1.72-1.74 (m, 0.6H), 1.87-1.90 (m, 0.4H), 3.69 (dd, J=12.02, 3.47Hz, 0.4H), 3.75 (dd, J=11.56, 6.24 Hz, 0.6H), 3.85 (dd, J=12.02, 3.47Hz, 0.6H), 3.92 (dd, J=11.56, 8.90 Hz, 0.4H), 7.18 (dd, J=8.55, 2.31 Hz,0.4H), 7.22 (dd, J=8.55, 2.31 Hz, 0.6H), 7.50 (d, J=2.31 Hz, 0.4H), 7.56(d, J=2.31 Hz, 0.6H), 7.59 (d, J=8.55 Hz, 0.6H), 7.60 (d, J=8.55 Hz,0.4H)

Step 43-{3-[(1R,3S)-1-(4-Bromo-3-chloro-phenyl)-2-hydroxymethyl-1,3-dimethyl-pentyl]ureido}bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester

3-(Methoxycarbonyl) bicyclo[1.1.1]pentane-1-carboxylic acid (5.2 g) andtoluene (50 mL) were mixed under argon, and thereto were addeddiphenylphosphoryl azide (7.2 mL) and triethylamine (4.6 mL) at roomtemperature. The reaction solution was stirred under heating at 110° C.for 1 hour. The reaction solution was added dropwise to a solution of(S)-2-[(R)-1-amino-1-(4-bromo-3-chloro-phenyl)-ethyl]-3-methyl-pentan-1-ol(9.8 g) in tetrahydrofuran (50 mL) over 10 minutes under ice cooling.The reaction solution was stirred at room temperature for 4 hours. Tothe reaction solution was added N,N,N′-trimethylethylenediamine (1.0ML), and the solution was concentrated under reduced pressure. To theresulting residue was added 10% aqueous citric acid solution, and themixture was extracted with ethyl acetate (twice). Combined organiclayers were washed with brine, and then dried over sodium sulfate.Sodium sulfate was removed through a filter, and then the filtrate wasconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography (ethyl acetate:n-hexane 1:1) to givethe title compound (10.4 g).

¹H-NMR (400 MHz, CDCl₃) 0.18 (d, J=6.94 Hz, 1.8H), 0.76 (d, J=6.94 Hz,1.2H), 0.80 (t, J=7.17 Hz, 1.2H), 0.87 (t, J=7.17 Hz, 1.8H), 1.23-1.28(m, 1H), 1.48-1.77 (m, 3H), 1.84 (s, 1.2H), 1.91 (s, 1.8H), 2.29 (s,2.4H), 2.29 (s, 3.6H), 3.67 (s, 3H), 3.67-3.70 (m, 0.6H), 3.77-3.85 (m,1H), 3.97 (dd, J=11.33, 8.32 Hz, 0.4H), 4.60 (brs, 1H), 7.09-7.16 (m,1H), 7.40-7.46 (m, 1H), 7.53 (d, J=8.32 Hz, 1H)

Step 53-[(S)-4-(4-Bromo-3-chloro-phenyl)-5-((S)-sec-butyl)-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl]-bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester

3-{3-[(1R,3S)-1-(4-Bromo-3-chloro-phenyl)-2-hydroxymethyl-1,3-dimethyl-pentyl]ureido}bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester (10.4 g) and dichloromethane (100 mL) were mixed underargon, and thereto were added (diacetoxyiodo)benzene (7.3 g) and2,2,6,6-tetramethylpiperidin-1-oxyl radical (0.323 g) at roomtemperature. The reaction solution was stirred at room temperature for16.5 hours. Then, thereto were added a mixed solution of sodiumthiosulfate (0.954 g) and water (40 mL) and saturated aqueous sodiumhydrogen carbonate solution under ice cooling. The layers of thereaction solution were separated, and the aqueous layer was extractedwith chloroform (twice). Combined organic layers were washed withsaturated aqueous sodium hydrogen carbonate solution and brine, and thendried over sodium sulfate. Sodium sulfate was removed through a filter,and then the filtrate was concentrated under reduced pressure. Theresulting residue was azeotroped with toluene. The resulting residue wasmixed in toluene (100 mL), and thereto was added pentafluoroanilinetrifluoromethanesulfonate (0.345 g) at room temperature. The reactionsolution was stirred under warming at 110° C. for 3 hours. The reactionsolution was concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography (ethylacetate:n-hexane=1:2) to give the title compound (4.6 g).

¹H-NMR (400 MHz, CDCl₃) 0.64 (d, J=6.70 Hz, 3H), 0.86 (t, J=7.28 Hz,3H), 1.25-1.36 (m, 1H), 1.40-1.51 (m, 1H), 1.53-1.63 (m, 1H), 1.67 (s,3H), 2.47 (s, 6H), 3.71 (s, 3H), 4.61 (s, 1H), 5.77 (s, 1H), 7.21 (dd,J=8.44, 2.31 Hz, 1H), 7.52 (d, J=2.31 Hz, 1H), 7.58 (d, J=8.44 Hz, 1H)

Step 63-{(S)-5-((S)-sec-Butyl)-4-[4-(2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester

3-[(S)-4-(4-Bromo-3-chloro-phenyl)-5-((S)-sec-butyl)-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl]-bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester (4.6 g), potassiumtrans-2-tert-butyl-cyclopropyl-trifluoroborate (2.4 g),[1,1′-bis(di-tert-butyl-phosphino)ferrocene]palladium (II) dichloride(0.62 g), cesium carbonate (9.4 g), toluene (50 mL), and water (5 mL)were mixed under argon. The reaction solution was stirred under heatingat 100° C. for 3.5 hours. To the reaction solution was added ammonium1-pyrrolidinecarbodithioate (0.69 g) under cooling with a water bath,and the mixture was stirred for 50 minutes. To the reaction solution wasadded additional ammonium 1-pyrrolidinecarbodithioate (0.69 g) at roomtemperature, and the mixture was stirred for 20 minutes. Insolublesubstances were filtered through Celite. The filtrate was separated, andthe resulting aqueous layer was extracted with ethyl acetate (twice).Combined organic layers were washed with brine, and then dried oversodium sulfate. Sodium sulfate was removed through a filter, and thenthe filtrate was concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography (ethylacetate:n-hexane=1:2) to give a diastereomer mixture (3.5 g). Theresulting diastereomer mixture (0.84 g) was purified with a chiralpreparative column to give the title compound (0.29 g).

Purification conditions for the preparative column are shown as follows.

Preparative apparatus: Recycling Preparative Liquid ChromatographLC-9225 NEXT SERIES, Japan Analytical Industry Co., Ltd.Column: Daicel CHIRALPAK IA 2.0 cmφ×25 cmMobile phase: hexane:2-propanol=93:7Flow rate: 10.0 mL/min

Detection: UV (220 nm)

Measurement with a chiral column showed 10.0 minutes of the retentiontime for the resulting title compound (9.4 minutes of the retention timefor diastereomers of the title compound) with 99% de purity. Analyticalconditions for the chiral column are shown as follows.

Measuring apparatus: HPLC system, Shimadzu Corporation,High-Performance Liquid Chromatograph Prominence Column: DaicelCHIRALPAK IA-30.46 cmφ×15 cmColumn temperature: 40° C.Mobile phase: hexane:2-propanol=95:5Flow rate: 1.0 mL/min

Detection: UV (220 nm)

¹H-NMR (400 MHz, CDCl₃) 0.63 (d, J=6.70 Hz, 3H), 0.83-0.77 (m, 1H), 0.85(t, J=7.28 Hz, 3H), 0.88-0.94 (m, 11H), 1.25-1.34 (m, 1H), 1.41-1.50 (m,1H), 1.58-1.63 (m, 1H), 1.66 (s, 3H), 2.08-2.12 (m, 1H), 2.47 (s, 6H),3.71 (s, 3H), 4.55 (s, 1H), 5.75 (s, 1H), 6.86 (d, J=8.09 Hz, 1H), 7.24(dd, J=8.09, 2.08 Hz, 1H), 7.37 (d, J=2.08 Hz, 1H)

Step 73-{(S)-5-((S)-sec-Butyl)-4-[4-((1R,2R)-2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid

3-((S)-5-((S)-sec-Butyl)-4-[4-(2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid methyl ester (0.287 g), tetrahydrofuran (2 mL), and methanol (2 mL)were mixed, and thereto was added 2N aqueous sodium hydroxide solution(0.58 mL) at room temperature. The reaction solution was stirred at roomtemperature for 18 hours. The reaction solution was concentrated, and tothe resulting residue were added 1N hydrochloric acid and water. Themixture was extracted with ethyl acetate (twice). Combined organiclayers were washed with brine, and then dried over sodium sulfate.Sodium sulfate was removed through a filter, and then the filtrate wasconcentrated under reduced pressure. To the resulting residue were addedacetonitrile (1.5 mL) and water (10 mL), and the mixture was stirred atroom temperature. The precipitated solid was filtered to give the titlecompound (0.268 g).

¹H-NMR (400 MHz, DMSO-D₆) 0.61 (d, J=6.70 Hz, 3H), 0.81 (t, J=7.40 Hz,3H), 0.86-0.91 (m, 12H), 1.22-1.32 (m, 1H), 1.39-1.49 (m, 1H), 1.56 (s,3H), 1.63-1.71 (m, 1H), 1.98-2.04 (m, 1H), 2.27 (s, 6H), 5.87 (s, 1H),6.98 (d, J=8.20 Hz, 1H), 7.04 (s, 1H), 7.20 (dd, J=8.21, 2.20 Hz, 1H),7.30 (d, J=2.20 Hz, 1H), 12.42 (brs, 1H)

Absolute configurations of the asymmetric carbons of the title compoundwere determined by single-crystal X-ray structural analysis.

Example 3 Crystalline polymorphs of3-{(S)-4-[4-((1R,2R)-2-tert-butyl-cyclopropyl)-3-chloro-phenyl]-5-isopropyl-4-methyl-2-oxo-3,4-dihydro-2H-pyrimidin-1-yl}bicyclo[1.1.1]pentane-1-carboxylicacid

X-ray powder diffraction (XRD), simultaneous measurement ofthermogravimetric analysis and differential thermal analysis (TG-DTA),and differential scanning calorimetry (DSC) were performed forCrystalline Forms A to E of the title compound. Measuring apparatus andconditions for each measurement are shown as follows.

Powder X-Ray Diffractometry:

Measuring apparatus: X'pert-PRO-MPD (Spectris Co., Ltd.)Measuring conditions: X ray, Cu/45 kV/40 mA, Analyzed by transmissionmethod

Simultaneous Measurement of Thermogravimetry Analysis and DifferentialThermal Analysis:

Measuring apparatus: TGA/SDTA851°/SF (Mettler Toledo International Inc.)Measuring conditions: Heating rate 5° C./min.

Differential Scanning Calorimetry:

Measuring apparatus: DSC Q2000 (TA Instruments Japan Inc.)Measuring conditions: Heating rate 10° C./min.

Each measurement result for each crystalline form is shown as below.

A. Crystalline Form A

XRD data is shown in FIG. 1. Diffraction angle 20 and diffractionintensity of each peak are shown as follows.

Pos. Relative intensity NET intensity [°2Th.] [%] [cts] 4.4150 88.60696.00 4.7253 92.34 725.34 9.3448 51.69 406.05 12.6233 89.18 700.5415.5239 54.31 426.65 15.9538 100.00 785.54 18.9756 93.38 733.57 19.380661.14 480.24 23.4055 52.66 413.64

DSC data is shown in FIG. 2. Enthalpy of endothermic peaks on the DSCcurve was 68.5 J/g, the endothermic temperature was 195° C., and theextrapolated onset temperature was 193° C.

B. Crystalline Form B

XRD data is shown in FIG. 3. Diffraction angle 20 and diffractionintensity of each peak are shown as follows.

Pos. Relative intensity NET intensity [°2Th.] [%] [cts] 7.3234 41.661172.05 8.6616 59.85 1683.77 10.3766 54.65 1537.60 11.3651 74.04 2083.0315.7159 100.00 2813.48 16.4562 33.94 954.94 17.0881 38.09 1071.7817.3618 97.60 2746.08 18.8969 91.19 2565.57 20.2651 27.35 769.36 21.293911.22 315.81 23.7512 11.98 336.95

DSC data is shown in FIG. 4. Enthalpy of endothermic peaks on the DSCcurve was 76.0 J/g, the endothermic temperature was 230.2° C., and theextrapolated onset temperature was 229.6° C.

C. Crystalline Form C

XRD data is shown in FIG. 5. Diffraction angle 20 and diffractionintensity of each peak are shown as follows.

Pos. Relative intensity NET intensity [°2Th.] [%] [cts] 7.4077 100.0018820.91 9.9304 14.05 2644.11 10.4586 9.36 1760.82 11.3577 6.59 1240.8111.5929 20.31 3821.92 13.3543 32.22 6063.80 14.1920 6.30 1186.13 17.410141.14 7743.09 18.2839 20.79 3912.93 18.6516 54.77 10307.31 19.3578 33.496302.58

DSC data is shown in FIG. 6.

D. Crystalline Form D

XRD data is shown in FIG. 7. Diffraction angle 2θ and diffractionintensity of each peak are shown as follows.

Pos. Relative intensity NET intensity [°2Th.] [%] [cts] 4.2615 100.001492.44 9.4092 32.36 482.89 14.1307 71.27 1063.67 14.9941 54.60 814.8916.0213 26.03 388.41 17.9068 24.55 366.47 18.6997 76.03 1134.65 20.223117.90 267.20 23.1278 16.36 244.14

TG-DTA data is shown in FIG. 8. The rate of weight decrease was 3.6% bydehydration. This value corresponded to the theoretical water content ofa monohydrate of the title compound, and Crystalline Form D was deemedto be a monohydrate of the title compound.

DSC data is shown in FIG. 9. Enthalpy of endothermic peaks on the DSCcurve was 118.4 J/g, the endothermic temperature was 128.0° C., and theextrapolated onset temperature was 118.9° C.

E. Crystalline Form E

XRD data is shown in FIG. 10. Diffraction angle 20 and diffractionintensity of each peak are shown as follows.

Pos. Relative intensity NET intensity [°2Th.] [%] [cts] 4.2178 100.001955.54 9.6770 17.62 344.56 13.7062 39.21 766.72 13.9822 39.97 781.7115.1994 21.16 413.75 15.4232 21.09 412.41 16.9120 24.01 469.52 18.809423.63 462.14 20.4669 15.16 296.44 21.9321 15.19 296.96 22.3640 9.64188.43

TG-DTA data is shown in FIG. 11. The rate of weight decrease was 3.9% bymelting. This value corresponded to the theoretical water content of amonohydrate of the title compound, and Crystalline Form E was deemed tobe a monohydrate of the title compound.

DSC data is shown in FIG. 12. Enthalpy of endothermic peaks on the DSCcurve was 138.1 J/g, the endothermic temperature was 125.9° C., and theextrapolated onset temperature was 115.0° C.

Test Example 1 In-Vitro Assay for Inhibitory Effect Against RORγTranscriptional Activity

Inhibitory effect against RORγ transcriptional activity was assessed byreporter gene assay for test compounds.

cDNAs encoding human and mouse RORγ ligand binding domains (LBD) wereobtained based on the sequences of human RORγ (Genbank registered numberNM_005060.3) and mouse RORγ (Genbank registered number NM_011281.2) (LBDsequences: from Ser253 to Lys518 for human RORγ; from Ile251 to Lys516for mouse RORγ).

LBD cDNAs of human and mouse RORγ were inserted into pFA-CMV vector(Agilent Technologies, Inc.), which expresses GAL4-DNA binding domainfusion protein. The resulting plasmids are hereinafter referred to aspFA/hRORγ plasmid and pFA/mRORγ plasmid, respectively.

pFA/hRORγ plasmid or pFA/mRORγ plasmid was transiently co-transfectedinto Chinese hamster ovary cells (CHO cells) with pG5-Luc (Promega),reporter plasmid expressing firefly luciferase in a GAL4-dependentmanner.

TransIT (Registered trademark) CHO Transfection Kit (Mirus) was used toco-transfect CHO cells with the plasmids. One day prior to the assay,CHO cells were suspended in HAM F-12 Nutrient medium containing 10%(v/v) fetal bovine serum and seeded at 5.5×10⁶ cells per 225 cm² cellculture flask each. 72 μL of TransIT (Registered trademark) CHO Reagentwas added into a 2 mL tube containing 1.55 mL of Opti-MEM, and thenmixed and incubated at room temperature for 10 min. 50.4 μL of a plasmidsolution containing 300 ng of pFA/hRORγ plasmid, 12000 ng of pG5-Lucplasmid, and 11700 ng of pcDNA3.1 plasmid were added into the tube andmixed gently. In the case of mouse assay, a plasmid solution containing300 ng of pFA/mRORγ plasmid, 12000 ng of pG5-Luc plasmid, and 11700 ngof pcDNA3.1 plasmid was added, instead. The mixture was incubated atroom temperature for 10 min. 12 μL of CHO Mojo Reagent was then addedinto each tube and mixed gently. The mixture was incubated at roomtemperature for 10 min. The resulting transfection reagent was appliedto the cell culture. After incubation at 37° C., 5% CO₂ for 4 hours, theplasmid-transfected CHO cells were harvested by trypsin treatment. Thecollected cells were resuspended in culture medium and plated into a384-well-white plate at 8,000 cells/35 μL/well. The plate was let standat room temperature for 1 hour and then incubated at 37° C., 5% CO₂ for3 hours. Test compounds were dissolved in dimethyl sulfoxide (DMSO) toobtain a concentration of 10 mM. Each solution was serially diluted withDMSO and further diluted with culture medium just before use. The testcompound solutions were added to the cells in the plate at 6 differentconcentrations. The final concentration of DMSO was 0.2% (v/v). Afterthe addition of test compound, the cells were incubated at 37° C., 5%CO₂ for 2 days.

Cell viability was tested by luminescence method with CellTiter-Glo(Promega). Two days after the addition of test compound, 40 μL each ofCellTiter-Glo was added into the 384-well-plate. Ten minutes after theaddition, luminescence was measured for each well with a microplatereader. The luminescence count in the cells treated with 0.2% DMSO alonewas defined as 100%, and the cell viability after treatment with thetest compound was calculated as a percentage (%-of-control) based on thevalue of 0.2% DMSO alone. When the cell viability was 70% or less, atest compound was assessed to have cytotoxicity.

RORγ transcriptional activity was detected as the intracellularluciferase activity using SteadyLite HTS Reporter Gene Assay System(Perkin Elmer). StedyLite Reagent was diluted 2.5-fold with Extensionbuffer (10 mM Tricine, 0.2% (w/v) bovine serum albumin, 0.02% (v/v)Tween-20) to obtain a luciferase substrate solution. Two days after theaddition of test compound, 40 μL each of the luciferase substratesolution was added into the 384-well-plate. After the incubation at roomtemperature for 10 minutes, luminescence of each well was measured witha microplate reader. The luciferase activity derived from theluminescence count in a vehicle-control well treated with 0.2% DMSOalone was defined as 100%, and the luciferase activity after treatmentwith the test compound was calculated as a percentage (%-of-control)based on the value of the vehicle-control. An EC₅₀ value of the testcompound was calculated by curve fitting. The luminescence counts at theconcentration of test compound where cytotoxicity was observed wereexcluded from data analysis.

The results are shown in the table below.

LUC EC₅₀ (μM) Compound hRORγ mRORγ (1) 0.023 0.023 (2) 0.014 0.029

Test Example 2 Assessment for In-Vitro Metabolic Stability A. MetabolicStability in Liver Microsomes (1) Preparation of a Test Solution

A test solution was prepared by diluting 10 mM DMSO solution of a testsubstance with acetonitrile by 100 times.

(2) Metabolic Stability Study with Liver Microsomes

Human and animal liver microsomes (SEKISUI XENOTECH; Human (H2610), Rat(R1000), Mouse (M1000), Monkey (P2000), Dog (D1000)) were adjusted withpotassium phosphate buffer (pH 7.4) to 0.2 mg/mL in the reaction system.Then, 1% of the solution of a test substance prepared was added to thereaction system. Thereto was added an NADPH generating system (preparedaccording to the method of Non Patent Literature 28), and the metabolicreaction was initiated. At a designated time, 0.1% formicacid-acetonitrile/water (3:1) was added thereto, and the reaction wasterminated.

(3) Analysis by LC/MS

The sample after termination of the reaction was separated bycentrifugation (1400 g, 20 min.), and then the supernatant was measuredby LC/MS (UPLC-SQD (waters)) to calculate a residual ratio of unreactedsubstances.

B. Metabolic Stability in Hepatocytes (1) Preparation of a Test Solution

A test solution was prepared by diluting 10 mM DMSO solution of a testsubstance with acetonitrile by 20 times.

(2) Metabolic Stability Study with Hepatocytes

Human and animal cryopreserved hepatocyte (BIOIVT; Human (IVT-X008000),Rat (IVT-M00005), Monkey (IVT-M00305), Dog (IVT-M00205)) were adjustedwith William's Medium E (SIGMA; W1878) to 1×10⁶ cells/mL of a cellsuspension, which was then added to a 96-well plate. Then, 1% of thesolution of a test substance prepared was added to the reaction system,and the metabolic reaction was initiated. At a designated time, 0.1%formic acid-acetonitrile was added thereto, and the reaction wasterminated.

(3) Analysis by LC/MS

The sample after termination of the reaction was separated bycentrifugation (1400 g, 20 min.), and then the supernatant was measuredby LC/MS (UPLC-SQD (waters)) to calculate a residual ratio of unreactedsubstances.

Results are shown in the following table.

Metabolic stability in liver microsomes at 60 min. (%) Compound HumanRat Mouse Monkey Dog (1) 99 103 95 93 98 (2) 96 95 98 96 95

Metabolic stability in hepatocytes at 240 min. (%) Compound Human RatMonkey Dog (1) 94 90 83 88 (2) 94 94 85 78

Test Example 3 Measurement of Potential for Induction of CYP3A4

Potential for induction of CYP3A4 was evaluated with Nuclear ReceptorActivation Kits (PURACYP; DPX2-96-002) according to protocols attached.

The following experiment was performed with substances accompanyingNuclear Receptor Activation Kits (PURACYP; DPX2-96-002).

A measurement method is specifically shown as follows. DPX2 cells wereseeded on a 96-well plate and incubated with a CO₂ incubator for 24hours. A compound-supplemented medium was prepared by adding 0.1% of aDMSO solution of test compound (1, 3, 10 mM) or rifampicin (10 mM) as apositive control to a medium. The media on 96-well plate were removedwith an aspirator 24 hours after seeding, and thereto was added thecompound-supplemented medium which was prepared. CellTiter-Fluor wasadded to the medium 48 hours after the addition of compound, and themixture was incubated with a CO₂ incubator for 1 hour. Then, a cellviability was measured by the fluorometric method. ONE-Glo was added tothe mixture, and the mixture was incubated with a CO₂ incubator for 5minutes. Then, the reporter activity was measured by luminescentdetermination. The induction ratio was calculated by comparing theresulting reporter activity with a DMSO control sample. % of positivecontrol was calculated from the following equation.

${\%\mspace{14mu}{positive}\mspace{14mu}{control}} = \frac{\begin{pmatrix}{{{activity}\mspace{14mu}{of}\mspace{14mu}{test}\mspace{14mu}{drug}\mspace{14mu}{treated}\mspace{14mu}{cells}} -} \\{{activity}\mspace{14mu}{of}\mspace{14mu}{negative}\mspace{14mu}{control}}\end{pmatrix} \times 100}{( {{{active}\mspace{14mu}{of}\mspace{14mu}{positive}\mspace{14mu}{control}} - {{activity}\mspace{14mu}{of}\mspace{14mu}{negative}\mspace{14mu}{control}}} )}$

Results are shown in the following table.

Potential for induction of CYP3A4 (%) Compound 3 μM 10 μM 30 μM (1) <10<10 <10 (2) <10 <10 ND ND: Not determined.

Test Example 4 Measurement of Solubility

A 10 mM DMSO solution (10 μL) of test substance was added to a 96-wellplate, and the mixture was concentrated and dried with a centrifugalevaporator (Genevac HT-4X). 200 μL of each solvent (JP1, JP2, FaSSIF,FeSSIF) (see Non Patent Literatures 29 and 30) was added to the wellafter evaporation. The mixture was shaken at room temperature at 2500rpm for 4 hours. Then, the mixture was filtered in two parts withMultiScreen PCF filter (MERCK Millipore; MSSLBPC) to give samples (50and 100 μL). 40 μL was collected from the sample obtained in the secondfiltration, and thereto was added acetonitrile (300 μL). Aftercentrifugation (4000 g, 5 min.) of the mixture, a part of thesupernatant was measured by LC-UV/MS (UPLC-Premier (waters)).

Results are shown in the following table.

Solubility (μM) Compound JP1 JP2 FaSSIF FeSSIF (1) <0.8 451 470 ≥475 (2)<0.7 447 445 463

Test Example 5 Rat Pharmacokinetic (PK) Assay and Measurement of PlasmaConcentrations (1) Rat PK Study

A DMSO solution (0.1 mL/kg) of test substance (0.3 mg/kg) wasadministered to a male rat (7-week old), and blood was collected atdesignated times (5, 10, 15, 30 min., 1, 2, 4, 8, 25 hr). Samples afterthe blood collection were separated by centrifugation (11000 g, 5 min.)to obtain plasmas.

(2) Measurement of Plasma Concentrations

A double volume of a mixed solution of acetonitrile/water (9:1) wasadded to a plasma sample obtained in (1) in order that proteins wereextracted from the mixture. The sample after extraction was separated bycentrifugation (11000 g, 5 min.), and the supernatant was measured byLC/MSMS (Nexera (Shimadzu)-QTrap5500 (AB SCIEX)). A calibration curvewas measured simultaneously, and plasma concentrations were calculated.

The elimination rate constant (Kel) was calculated from two points ofthe elimination phase in the plasma concentration profile, and thehalf-life (T1/2) was calculated from the elimination rate constant.

Kel=−(ln Conc1−ln Conc2)/(t1−t2)

T1/2=0.693/Kel

Results are shown in the following table.

Compound Plasma half-life (h) (1) 10.5 (2) 13.2

Test Example 6 Effects of Test Substances on IL-17 Production in Plasmasof Antigen-Sensitized Mice

Effects of test substances on IL-17 production in plasmas were assessedin antigen-sensitized mice. Antigen sensitization was performedaccording to Non Patent Literature 31. Nine-week old female C57BL/6Jmice (Japan Charles River K. K.) were used as a test animal.

An emulsion prepared by mixing 3 mg/mL of myelin oligodendrocyteglycoprotein (MOG; ANASPEC) and 4 mg/mL of complete Freund's adjuvant(CFA; Chondrex) in an equal ratio was administered subcutaneously to amouse at the bilateral abdomens at a volume of 50 μL/site (day 1). 1μg/mL of pertussis toxin (PTX; List Biological Laboratories) wasadministered intraperitoneally at a volume of 200 μL/head (day 1 and 3).Blood was collected at day 8.

Administration was performed once daily for 7 days after theadministration of emulsion (day 1 to 7). 0.5% (w/v) methyl cellulose(MC) was administered orally at a volume of 10 mL/kg to Normal group(normal mice) and Vehicle group (antigen-sensitized mice), and 0.03mg/mL or 0.1 mg/mL of a test substance suspended in 0.5% (w/v) MC wasadministered orally at a volume of 10 mL/kg to a compound-administeredgroup.

Blood was separated by centrifugation, and then the IL-17 concentrationin plasma was measured with Quantikine (Registered trade mark) MouseIL-17 ELISA Kit (R&D systems, Inc.).

The IL-17 concentration of Normal group was deemed to be 0% and that ofVehicle group was deemed to be 100%. The IL-17 concentration percentage(% of control) of the compound-administered group was calculated fromthe following equation.

% of control=(a−b)/(c−b)×100

a: IL-17 concentration of the compound-administered groupb: IL-17 concentration of the Normal groupc: IL-17 concentration of the Vehicle group

Then, a dose of compound where the IL-17 concentration was decreased by50% (ED₅₀) was calculated from the following equation.

ED₅₀=10 ^((log(A/B)×(50−D)/(C−D)+log(B)); provided that ED₅₀=B in thecase D=50, and ED₅₀>B in the case D>50A: dosed amount of the lower dose group (i.e. 0.3 mg/kg)B: dosed amount of the higher dose group (i.e. 1 mg/kg)C: % of control at the lower dose groupD: % of control at the higher dose group

Results are shown in the following table.

Compound Dose (mg/kg) % of control ED₅₀ (mg/kg) (1) 0.3 (=A) 66 (=C) 0.51 (=B) 24 (=D) (2) 0.3 (=A) 85 (=C) >1 1 (=B) 54 (=D)

Test Example 7 Effects of Test Substances on IL-17 Production in Plasmasof Cytokine-Stimulated Normal Mice

Effects of test substances on IL-17 production in plasmas were assessedin normal mice. The assessment was performed according to Non PatentLiterature 32. Nine-week old female C57BL/6J mice (Japan Charles RiverK. K.) were used as a test animal.

A mixed solution prepared by mixing Recombinant Mouse IL-1 beta/IL-1F2Protein (R&D systems, INC.) and Recombinant Mouse IL-23 Protein (R&Dsystems, INC.) at a final concentration of 1.5 μg/mL was administeredintravenously to all groups at the tail vein at a volume of 200 μL/head.Blood was collected 2 hours after the administration of cytokinesolution.

Administration was performed 28 hours before the administration ofcytokine solution. 0.5% (w/v) methyl cellulose (MC) and 0.1 mg/mL of atest substance suspended in 0.5% (w/v) MC were administered orally at asingle volume of 0.2 mL/head to Vehicle group and acompound-administered group, respectively.

Blood was separated by centrifugation, and then the IL-17 concentrationin plasma was measured with Quantikine (Registered trade mark) MouseIL-17 ELISA Kit (R&D systems, Inc.).

The IL-17 concentration of Vehicle group was deemed to be 100%, and theIL-17 concentration percentage of the compound-administered group to theVehicle group was calculated from the following equation. Results areshown in the following table.

%=a/b×100a: IL-17 concentration of the compound-administered groupb: IL-17 concentration of the Vehicle group

Compound IL-17 concentration (%) (1) 52 (2) 67

Formulation Examples

Formulation Examples in the present invention include, for example, thefollowing formulations. The present invention, however, is not intendedto be limited to these Formulation Examples.

Formulation Example 1. (Preparation of a Capsule

1) Example 1 Compound 30 mg 2) Microcrystalline cellulose 10 mg 3)Lactose 19 mg 4) Magnesium stearate 1 mg

Ingredients 1), 2), 3), and 4) are mixed to be filled in a gelatincapsule.

Formulation Example 2 (Preparation of a Tablet

1) Example 1 Compound 10 g 2) Lactose 50 g 3) Cornstarch 15 g 4)Carmellose calcium 44 g 5) Magnesium stearate 1 g

The total amount of Ingredients 1), 2), and 3) and 30 g of Ingredient 4)are combined with water, dried in vacuo, and then granulated. Theresulted granules are mixed with 14 g of Ingredient 4) and 1 g ofIngredient 5), and tableted with a tableting machine. In this manner,1000 tablets comprising 10 mg of Example 1 Compound per each areobtained.

INDUSTRIAL APPLICABILITY

A compound of Formula (1) or (2) or a pharmaceutically acceptable saltthereof is expected to be useful for treating or preventing autoimmunediseases, allergic diseases, dry eye, fibrosis, cancers, metabolicdisease, ischemia, cardiomyopathy, hypertension, and periodontaldisease.

1-12. (canceled)
 13. A method of preparing a compound of Formula (1):

the method comprising reacting a compound of the following formula:

with sodium hydroxide.
 14. The method of claim 13, wherein the reactingis carried out at room temperature in the presence of isopropanol.
 15. Amethod of preparing a compound of the following formula:

the method comprising reacting a compound of the following formula:

with (diacetoxyiodo)benzene in the presence of2,2,6,6-tetramethylpiperidine-1-oxyl radical.
 16. The method of claim15, wherein the reacting is carried out at room temperature in thepresence of acetic acid and cyclopentyl methyl ester.
 17. A compoundselected from the group consisting of:


18. The compound of claim 17, wherein the compound has the formula:


19. The compound of claim 17, wherein the compound has the formula:


20. The compound of claim 17, wherein the compound has the formula:


21. The compound of claim 17, wherein the compound has the formula:


22. The compound of claim 17, wherein the compound has the formula:


23. The compound of claim 17, wherein the compound has the formula:


24. The compound of claim 17, wherein the compound has the formula:


25. The compound of claim 17, wherein the compound has the formula:


26. The compound of claim 17, wherein the compound has the formula:


27. The compound of claim 17, wherein the compound has the formula:


28. The compound of claim 17, wherein the compound has the formula:


29. The compound of claim 17, wherein the compound has the formula:


30. A compound selected from the group consisting of:


31. The compound of claim 30, wherein the compound has the formula:


32. The compound of claim 30, wherein the compound has the formula:


33. The compound of claim 30, wherein the compound has the formula:


34. The compound of claim 30, wherein the compound has the formula:


35. The compound of claim 30, wherein the compound has the formula:


36. The compound of claim 30, wherein the compound has the formula:


37. A crystalline form of a compound of Formula (1):

showing a powder X-ray diffraction pattern having three or more peaksselected from the group consisting of 7.4±0.2°, 9.9±0.2°, 10.5±0.2°,11.4±0.2°, 11.6±0.2°, 13.4±0.2°, 14.2±0.2°, 17.4±0.2°, 18.3±0.2°,18.7±0.2°, and 19.4±0.2° of the diffraction angle (2θ) measured withCuKα radiation.